Biology subjects

Jones, M.

Publications and source records attributed to Jones, M..

At least 19 recordsLinked to original sources

bioRxiv2026

Ectopic expression of the germline transcription factor LSL-1 contributes to developmental delay following failed maternal epigenetic reprogramming

Proper transmission of cell identity between generations requires maternal epigenetic reprogramming mechanisms that prevent inappropriate inheritance of lineage-specific transcriptional programs. In Caenorhabditis elegans, loss of the H3K4me1/2 demethylase SPR-5 and the H3K9 methyltransferase MET-2 results in ectopic expression of germline genes in somatic tissues and severe developmental delay. Previous studies demonstrated that the chromatin regulator MES-4 contributes to these defects, but whether germline-specific transcription factors also participate in the ectopic transcriptional program remained unclear. Here, we investigated the role of the germline transcription factor LSL-1 in animals lacking SPR-5 and MET-2. We found that genes normally regulated by LSL-1 in the germline are significantly overrepresented among genes ectopically expressed in the soma of spr-5; met-2 progeny. Consistent with this observation, an endogenously tagged LSL-1 protein became ectopically expressed throughout somatic tissues when maternal SPR-5 and MET-2 activity was disrupted. Furthermore, depletion of LSL-1 partially suppressed the developmental delay observed in spr-5; met-2 mutants. Transcriptomic analyses revealed extensive overlap between MES-4- and LSL-1-dependent transcriptional programs, with most LSL-1-dependent genes also requiring MES-4. Notably, ectopic expression of lsl-1 itself depended on MES-4, suggesting that LSL-1 functions downstream of MES-4. Genes dependent on LSL-1 were strongly enriched for germline-associated expression programs and included previously identified direct LSL-1 targets. Together, our findings support a model in which MES-4 promotes ectopic expression of LSL-1, which in turn contributes to a shared germline-associated transcriptional program and developmental delay following failed maternal epigenetic reprogramming. These results demonstrate how lineage-restricted transcription factors cooperate with inherited chromatin states to reinforce aberrant transcriptional programs and disrupt cell fate boundaries.

developmental biology↗

bioRxiv2026

Beyond the Panglossian paradigm: adaptation, constraint, and anuran functional trait evolution.

Recognizing patterns in functional trait evolution is a necessary step in testing macroevolutionary questions. Quantification of these patterns and interpretation of their generative processes relies on an ever-expanding suite of comparative approaches, but current methods oversimplify the process-to-pattern mapping. This simplification may promote binary classifications of patterns and their drivers, such as adaptive versus non-adaptive or constrained versus unconstrained. A potentially more robust and evolutionarily informative alternative is to fit an expanded suite of evolutionary models at different levels of taxonomic or ecological resolution to reveal how evolutionary drivers imprint simultaneously on observed patterns of trait evolution. Here, we perform a clade-wide analysis of functional trait evolution across Anura (frogs and toads). Focusing on three functionally important traits, we quantify the relative fit of random, directional, and bounded models at the intra- and inter-microhabitat levels to explore how changes in trait function across microhabitats translate into different evolutionary regimes. We recover heterogeneous regime-level distributions of model support that imply complex underlying evolutionary dynamics, while also revealing methodological biases and model identifiability issues. These findings underscore the need to develop more robust tools for evolutionary model fitting and advance beyond binary frameworks for interpreting evolutionary processes.

evolutionary biology↗

bioRxiv2026

Introducing PHJ Media: A Unique Machine Learning -Driven Basal Formulation to Overcome Recalcitrance for Multi-Genotype Micropropagation of Cannabis sativa L.

Applications of tissue culture are critical for Cannabis sativa L. (cannabis), supporting clonal propagation, germplasm preservation, pathogen elimination, among other biotechnological applications. However, extensive genetic diversity associated with cannabis results in highly variable responses to in vitro conditioning, and no consensus basal media formulation exists to support reproducible micropropagation across genotypes. To address these limitations, a hybridized ensemble-NSGA-II approach was employed for concurrent optimization of individual media components to create a species specific, cultivar inclusive basal salt formulation for cannabis micropropagation. The resulting PHJ media represents a unique formulation that overcomes recalcitrance across a wide array of cannabis cultivars, facilitating improved growth and uniformity for the nine cultivars used in its development and validation. These results remain consistent from explant initiation through multiple rounds of subculture. The ability of PHJ to overcome genotypic recalcitrance is telling of its potential applicability with an array of plant species beyond cannabis. Additionally, robust performance both with and without plant growth regulators underscores the plausible use of PHJ for diverse applications beyond standard micropropagation. Ultimately, this cultivar-inclusive basal medium demonstrates utility for both scientific research and industrial-scale operations.

plant biology↗

bioRxiv2026

Periplasmic detoxification of urate hydroperoxide underpins E. coli survival in the inflamed gut

Uric acid, the final product of purine metabolism in humans, accumulates in blood and tissues at relatively high concentrations1 as humans lack the enzyme uricase2,3. Under inflammatory conditions, uric acid can be oxidised to yield reactive intermediates4. In activated neutrophils, myeloperoxidase (MPO) catalyses the oxidation of uric acid by hydrogen peroxide, leading to the formation of urate hydroperoxide (UH)5,6. While recent studies have shown that UH is toxic to bacteria lacking peroxiredoxins7, its precise mechanism of toxicity and the existence of dedicated bacterial defence systems remain unknown. Here, we identify HiuH as a periplasmic enzyme, conserved across E. coli strains, that specifically degrades UH. Our findings reveal that UH selectively induces the expression of hiuH and that HiuH efficiently detoxifies UH both in vitro and in bacterial cells. HiuH cooperates with MsrP, a periplasmic methionine sulfoxide reductase that repairs UH-induced protein-bound methionine oxidation. This combined defence offering both direct detoxification and damage repair, is essential for bacterial survival under UH stress, and confers a competitive fitness advantage in a DSS-induced mouse model of colitis. Although UH is chemically transient, our work shows that it imposes durable biological consequences and a sufficient fitness cost in the in vivo niches occupied by E. coli to favour the evolution of a dedicated detoxification pathway beyond general oxidative-stress responses, defining a key adaptation to periods of gut inflammation.

microbiology↗

bioRxiv2026

Mapping fibrotic microenvironments: single-cell and spatial profiling of Schistosoma mansoni-induced tissue fibrosis

Schistosoma-induced tissue fibrosis, driven by immune response to trapped schistosome eggs, is the principal cause of pathology and schistosomiasis-related morbidity. While praziquantel effectively clears adult parasites, no therapies exist to target eggs or egg-induced tissue fibrosis. Here, we utilise single-cell spatial transcriptomics to define the high-resolution molecular architecture of the fibrotic niche in Schistosoma mansoni-infected mice at 8 weeks post-infection. Our data reveal that the host executes divergent, organ-specific biological programs: constructing a rigid, concentric fibrotic granuloma for permanent egg sequestration in the liver while building a flexible, discontinuous collagen architecture in the intestine to facilitate egg transit. Spatially resolved cell-cell interaction analysis, unprecedented in schistosome-induced fibrosis, identifies the strongest cellular interactions between macrophages and collagen-producing cells, with TGF-{beta}-dominant ligand-receptor pairs in the hepatic niche and integrin-centered pairs in the intestine. Despite this architectural divergence, we detected a core signature of five ligand-receptor pairs shared across both tissues, suggesting shared signalling interactions that could be leveraged for pan-tissue therapeutics for treating schistosome-induced fibrosis. Together, these findings provide the first high-resolution spatial atlas of schistosomiasis-associated fibrosis and provide a foundational framework for discovering novel drug targets capable of reversing established tissue damage, addressing a critical clinical gap where current therapies fail.

molecular biology↗

bioRxiv2026

Unlocking a flexible set of phylogenetic models for discrete and continuous trait evolution using discretized stochastic diffusion

The practical utility of many modern phylogenetic comparative methods can depend on how accurately mathematical models capture the evolutionary process of traits. Boucher and Demery (2016) described a new quantitative trait model for testing hypotheses about constraint on phenotypic character evolution: Brownian motion with reflective limits. Since their analytic solution for the probability function under this bounded scenario was intractable for reasonably-sized trees, Boucher and Demery (2016) also identified a creative technique for computing the likelihood of their model. The basis of this method derives from the convergence of an equal-rates, symmetric, ordered Markov chain and continuous stochastic diffusion in the limit as the number of lattice states in the chain goes to{infty} (or, alternatively, as their widths decrease towards zero). We realized that this general approach had the potential to unlock a surprisingly large number of additional models for the phylogenetic comparative analysis of discrete and continuous trait data, and we explore several of these in the present article. Specifically, we examine application of this discretized diffusion approximation to the threshold model from evolutionary quantitative genetics, to a new "semi-threshold" trait evolution model, to a joint model where the rate of evolution for a continuous trait depends on the value of a co-evolving discrete character, along with a separate model where precisely the converse is true, and to a discrete-character-dependent multi-trend trended continuous trait evolution model. We conclude with some context for the origins of our article and discussion of other possible applications of this powerful approach.

evolutionary biology↗

bioRxiv2026

Early enhanced control of Plasmodium yoelii infection in IL-10-deficient mice is independent of IFN-γ, IL-12, and the humoral response

The outcome of a Plasmodium infection depends on the timely regulation of the robust pro-inflammatory response required to eliminate the parasite, but this response can cause tissue damage if not properly controlled. IL-10 is an important regulatory cytokine that prevents immunopathology during many Plasmodium infections; however, this protection comes at the expense of less effective parasite control. This is illustrated by infection with P. yoelii, in which mice exhibit a lower parasite load in the absence of IL-10. However, the immune components that limit parasite burden in the absence of IL-10 remain poorly understood. Abolishing IL-10 led to a predicted increase in TH1 polarization and higher production of IL-12 and IFN-{gamma}. However, the enhanced production of these cytokines did not explain the improved parasite control seen in Il10-/- mice. Loss of IL-10 signaling reduced the accumulation of germinal center B cells and plasmablasts in the spleen, indicating a role for IL-10 in supporting the humoral response. However, although B cells are essential for survival, they do not play a critical role in early parasite control in IL-10-deficient mice. Moreover, Il10-/-mice lacking IFN-{gamma} and B cells can limit early parasite expansion, suggesting that IL-10 suppresses host-protective pathways beyond the functions of B cells and IFN-{gamma} in parasite control.

immunology↗

bioRxiv2026

Tenascin N contributes to spinal motor nerve morphogenesis during development

Spinal motor nerves are an integral component of the nervous system whose development requires the coordination of many diverse cell types, including motor neurons, glia, and muscle. Although several molecular mechanisms guiding these interactions are known, many remain to be uncovered. Extracellular matrix (ECM) proteins also play a critical role in motor nerve assembly, yet their functions are less understood compared to classical pathfinding and guidance cues. Here, we identify a role for tenascin-n (tnn), an ECM glycoprotein, in spinal motor nerve development in zebrafish. Using in situ hybridization and immunohistochemistry, we show that tnn/Tnn is expressed and localized along vertical myosepta and the border of the ventral neural tube during spinal motor nerve development. To assess its function, we generated a CRISPR/Cas9 mutant allele, tnnuva96, and performed in vivo imaging and morphological analysis throughout motor nerve development. Loss of tnn leads to a subtle and transient increase in ectopic motor axon exit and aberrant motor axon branching in the zebrafish trunk. Our findings reveal a previously unrecognized role for tnn in spinal motor nerve assembly and expand our understanding of the diverse molecular contributors to spinal motor nerve development and morphogenesis.

developmental biology↗

bioRxiv2025

In Situ Photoactivated Hydrogel Adhesive Dressings for Post Colon Polypectomies (PolypCures)

Colon polypectomy is a widely performed endoscopic procedure that reduces the incidence of colorectal cancer but leaves exposed colonic wounds susceptible to bleeding, perforation, and infection. The current standard of wound care, mechanical clips, is limited by technical complexity, high cost, and poor efficacy for large (> 2 cm in diameter) or difficult-to-access lesions. Here, we present PolypCure, an in situ photoactivated hydrogel adhesive dressing delivered via a single catheter, through either oozing or spraying methods, to achieve rapid, spatiotemporally controlled sealing and hemostasis (within 2 min) of colon lesions using the white light source integrated into standard endoscopic instruments. The hydrogel, composed of norbornene- and thiol-functionalized polyethylene glycol, carboxymethyl cellulose, and Eosin Y, exhibits mechanical properties comparable to colon tissue (G [~] 6 kPa), strong shear strength (> 15 kPa), low swelling (< 200%), and small pore size ([~] 20 {micro}m), ensuring long-term stability (up to 30 days) and protection against bacterial infiltration. The formulation also demonstrates biocompatibility and hemocompatibility. In an in vivo pig colon endoscopic mucosal resection (EMR) model, PolypCure fully adheres to large lesions for at least 3 days and promotes early-stage wound healing. Overall, PolypCure is a promising solution to wound management post-polypectomy.

bioengineering↗

bioRxiv2025

Large-scale culturing of the tree microbiome enables targeted disease suppression

The tree microbiome is essential for host health and pathogen suppression. Synthetic microbial communities (SynComs) are emerging as important tools to understand microbiome dynamics and engineer microbiomes to harness beneficial properties. However, while the rational design, assembly and application of SynComs requires representative microbiota isolate collections combined with functional information, microbial culture collections from tree species such as oak (Quercus) are critically lacking. Here, we generated an oak microbiota culture collection comprising >30,000 isolates from 150 oak trees across Britain, belonging to key bacterial and fungal taxa that represented 61% of the total bacterial sequences and 87% of total fungal sequences as determined by culture-independent sequencing. Over 22,000 isolates were screened for suppression of bacterial species associated with degradation of live stem tissue in trees affected by Acute Oak Decline (AOD), identifying 341 bacterial isolates that suppressed oak pathogens. In vitro screening of 40 randomly assembled SynComs demonstrated that oak microbiota SynComs can suppress oak pathogenic bacteria associated with AOD. Inoculation of a disease-suppressive SynCom into the stem of oak seedlings and logs prior to pathogen challenge reduced the quantities of the bacteria, Brenneria goodwinii and Gibbsiella quercinecans, by 56% and 87%, respectively, in seedlings, and 71% and 95% in logs. This work demonstrates that the tree microbiome can be engineered using disease suppressive SynComs.

microbiology↗

bioRxiv2025

Collagen Prolyl Hydroxylases Regulate HIF-α Levels Independently of pVHL in ccRCC

The hypoxia-inducible factors, HIF-1 and HIF-2, are master regulators of the hypoxia response. Under ambient conditions, both are hydroxylated by the HIF prolyl hydroxylases (HIF PHDs) resulting in ubiquitination by the pVHL E3 ligase complex, leading to subsequent proteasomal degradation. During hypoxia, the HIF PHDs are inhibited, resulting in HIF- stabilization and transcriptional activation of genes involved in the adaptation to hypoxia. Previous studies have shown that the collagen PHD, P4HA1, which promotes proline hydroxylation of collagen, inhibits the HIF PHDs by modulating levels of -ketoglutarate and succinate, thus enhancing HIF-1 stability by preventing pVHL-mediate degradation. Here, we investigate the role of collagen PHDs in the regulation of HIF-1/2 in the setting of pVHL deficiency in ccRCC. We show that the collagen PHDs P4HA1 and P4HA2 are required for HIF-1 translation and HIF-2 transcription and translation independently of pVHL function, through a mechanism regulated in part by P4HA1/2-driven collagen production. Thus, we reveal a novel pVHL-independent mechanism of HIF-1/2 regulation driven by P4HA1/2 in ccRCC. Since the HIFs have been strongly implicated in ccRCC initiation and progression, our data suggests that inhibition of P4HA1/2 may be a promising therapeutic strategy in ccRCC.

cancer biology↗

bioRxiv2025

Systematic molecular glue drug discovery with a high-throughput effector protein remodeling platform

Realising the promise of new medicines that operate through a targeted molecular glue-induced degradation mechanism requires systematic tools that can uncover the relevant principles of neomorphic protein-protein interactions. Whilst some monovalent glue degraders have been found through serendipity, the rules for small molecule attributes and the pairs or complexes of proteins that are amenable to drug-induced proximity control remain poorly articulated. Here we introduce a new approach to address this by using programmed libraries of intramolecularly edited proteins to expand protein surface landscapes and trigger new druggable interactions. We show that effector proteins, such as the E3 ligase Cereblon, can be engineered to provoke neomorphic activity by inducing the degradation of new client proteins and that these de novo interactions provide a blueprint from which new small molecule degraders can be built. As a demonstration of the approach, we use the platform to identify new non-IMiD molecular glue degraders of the oncology target GSPT1. SUMMARYO_LIMolecular glues are a highly important and promising new form of therapeutic agent, but rationalising their discovery has so far been impossible C_LIO_LIGlueSEEKER screening enables prospective monovalent drug discovery by using high-throughput deep mutational scanning to re-engineer the function of effector proteins like E3 ligases C_LIO_LIWe used this approach to enable the computational discovery of small molecule glues which degrade the oncology target GSPT1 and show how the technology can be used across new targets C_LI

bioengineering↗

bioRxiv2025

Multiplexed proximity labelling proteomics identifies a non-canonical Bcl-2 family interactome associated with apoptotic priming

Bcl-2 family proteins govern the intrinsic apoptotic pathway by regulating mitochondrial outer membrane permeabilisation (MOMP), releasing apoptogenic factors into the cytosol. How close a cell is to MOMP, termed mitochondrial priming, is determined by the interactions between different Bcl-2 proteins at the outer mitochondrial membrane (OMM). However, Bcl-2 proteins can also drive diverse processes that do not result in cell death, such as incomplete MOMP, sublethal caspase activation, pro-inflammatory signalling and regulation of cellular metabolism. To understand the wider Bcl-2 family interactome and how it might be involved in these processes, we undertook an unbiased proteomic BioID screen, using both pro- and anti-apoptotic Bcl-2 family members as bait proteins. We found that most high-confidence potential interactions in non-apoptotic cells were outside the canonical Bcl-2 family interactome. Analysis of how this interactome changed in response to either BH3-mimetics or full-length BH3-only proteins revealed dynamic changes at multiple organelles and membrane contact sites in response to altered mitochondrial priming. These findings underscore the complex and varied interactions of the Bcl-2 family proteins, expanding their scope and function beyond the frequently studies intra-family interactions.

cell biology↗

bioRxiv2025

A lipid metabolism defect is an underlying contributor to Diamond Blackfan anemia syndrome

Analysis of neither Diamond Blackfan anemia syndrome (DBAS) cohorts nor animal models has revealed a potential mechanism for the variable anemia phenotype, a key feature of this disease. Here, we utilized an established Rpl5Skax23-Jus/+ murine DBAS model in order to study this dynamic erythropoiesis deficiency. These haploinsufficient mice exhibit variably penetrant craniofacial and cardiac defects mimicking the phenotypes of DBAS patients bearing RPL5 mutations. We additionally discovered that this specific heterozygous splicing mutation is pathogenic and leads to partial intron retention. By examining the transcriptome of fetal liver erythroid progenitors at E12.5, we demonstrate that the downregulation of erythroid differentiation pathways is consistent with the DBAS phenotype. We also identified dysregulated transcription of lipid metabolism genes with significant reduction in the abundance of Scd1 in a subset of E12.5 mutant embryos at risk for erythroid failure. SCD1, a key enzyme that converts saturated to monounsaturated fatty acids, has not been previously linked to erythropoiesis or DBAS. When anemia was induced in adult mice, pretreatment with an SCD1 inhibitor resulted in improved erythropoiesis. This analysis suggests a key role of lipid metabolism in the variable anemia penetrance in DBAS and highlights a previously unappreciated pathway that may serve as a potential target for drug development. Key PointsO_LIThe variable anemia in Rpl5Skax23-Jus/+mice is triggered by intrinsic/extrinsic stress C_LIO_LIRpl5 haploinsufficient murine and human erythroid progenitors exhibit a lipid metabolism signature with downregulation of Scd1/SCD. C_LI

developmental biology↗

bioRxiv2025

Specialised RNA decay fine-tunes monogenic antigen expression in African trypanosomes

Antigenic variation is a sophisticated immune evasion strategy employed by many pathogens. Trypanosoma brucei expresses a single Variant-Surface-Glycoprotein (VSG) from a large genetic repertoire, which they periodically switch throughout an infection. Co-transcribed with the active-VSG within a specialised nuclear body are expression-site-associated-genes (ESAGs), involved in important host-parasite interactions, including protecting the parasite from human serum lytic effects, modulating the hosts innate immune response and uptake of essential nutrients. Despite expression within the same polycistron, there is a significant differential expression between ESAGs and VSGs (>140-fold), however, the regulatory mechanism has remained elusive for decades. Here, using a combination of genetic tools, super resolution microscopy, proteomics and transcriptomics analyses, we identified three novel proteins, which are recruited in a hierarchical manner, forming discreet sub-nuclear condensates that are developmentally regulated and negatively regulate ESAG transcripts. Among them, Expression-Site-Body-specific-protein-2 (ESB2) contains a nuclease domain that shares structural similarity to the endonuclease domain found in SMG6, a critical component of nonsense mediated decay in mammals. Mutation of key residues required for the nuclease activity impaired ESB2 localisation and function. Overall, our findings reveal a novel mechanism of post-transcriptional regulation and shed light on how specialised RNA decay can regulate expression of specific genes.

molecular biology↗

bioRxiv2025

Multiscale Entropy of Resting-State fMRI Signals Reveals Differences in Brain Complexity in Autism

BackgroundAtypical intrinsic brain activity has been widely observed in autism spectrum disorder (ASD), yet the temporal complexity of these neural signals remains underexplored. This study aimed to characterise differences in resting-state brain signal complexity between individuals with ASD and neurotypical controls using multiscale entropy (MSE). MethodsResting-state fMRI data were obtained from the Autism Brain Imaging Data Exchange I (ABIDE I), a large multi-site dataset including 397 participants (179 with ASC, 218 neurotypical controls; ASD: Mean age 16.43 years old, SD 7.17 years old; CON: Mean age 15.75 years old, SD 5.67 years old). Voxel-wise multiscale entropy (MSE) features were extracted across multiple temporal scales. Group comparisons were conducted using voxel-wise t-tests and mixed-effects models to identify region- and scale-specific alterations in brain signal complexity. ResultsIndividuals with ASD showed reduced MSE in prefrontal regions at coarser time scales and elevated MSE in posterior midline regions, including the posterior cingulate cortex and precuneus, at finer scales, followed by a decline across coarser scales. This pattern suggests a shift toward uncorrelated randomness in posterior regions and reduced long-range complexity in frontal areas. No significant associations were found between MSE features and ADOS scores. ConclusionsThese findings reveal spatially and temporally specific alterations in brain signal complexity in ASD, particularly within the default mode network. Multiscale entropy provides a complementary approach to traditional connectivity and single-scale entropy analyses, offering novel insights into the organisation of intrinsic brain activity in neurodevelopmental conditions.

neuroscience↗

bioRxiv2025

The endoplasmic reticulum promotes microtubule organization and region-specific disassembly to execute Compartmentalized Cell Elimination

Specialized cells, such as neurons, die during development and disease. How subcellular organization and interactions across diverse compartments direct death is not well-understood. We examine this by studying the Compartmentalized Cell Elimination (CCE) developmental death program of the C. elegans tail-scaffolding cell (TSC). We find endoplasmic reticulum (ER) shape genes and the microtubule (MT) severase SPAS-1/Spastin, all linked to neurodegeneration, cooperate to promote CCE. Super-resolution imaging reveals profound spatiotemporal dynamics of MTs and the ER across CCE, including enrichment in a stereotyped degenerative node. We observe an ER-dependent non-centrosomal microtubule organizing center (ncMTOC) in the degenerative node, where the ER locally promotes both MT organization and SPAS-1/Spastin-mediated MT severing. The ER is spatially confined, such that SPAS-1/Spastin also has an ER-independent role. Our study expands our understanding of the cell biology of specialized cell death during development and presents molecular links to pathological neurodegeneration, paving the way to neurodegenerative therapies.

cell biology↗

bioRxiv2025

Identifiability of Phylogenetic Level-2 Networks under the Jukes-Cantor Model

AO_SCPLOWBSTRACTC_SCPLOWWe investigate which evolutionary histories can potentially be reconstructed from sufficiently long DNA sequences by studying the identifiability of phylogenetic networks from sequence data generated under site independent models of molecular evolution. While previous work in the field has established the identifiability of phylogenetic trees and level-1 networks, networks with non-overlapping reticulation cycles, less is known about more complex network structures. In this work, we extend identifiability results to network classes that include pairs of tangled reticulations. Our main result shows that binary semi-directed level-2 phylogenetic networks are generically identifiable under the Jukes-Cantor model, provided they are triangle-free and strongly tree-child. We also strengthen existing identifiability results for level-1 networks, showing that the number of reticulation nodes is generically identifiable under the Jukes-Cantor model. In addition, we present more general identifiability results that do not restrict the network level at all and hold for the Jukes-Cantor as well as for the Kimura-2-parameter model. Specifically, we demonstrate that any two binary semi-directed networks that display different sets of 4-leaf subtrees (quartets) are distinguishable. This has direct implications for the identifiability of a networks reticulated components (blobs). We show that the tree-of-blobs of a network, the global branching structure of the network, is identifiable, as well as the circular ordering of the subnetworks around each blob, for networks in which edges do not cross and taxa are on the outside.

evolutionary biology↗