Mathien, S. | Search bioRxiv

bioRxiv2025

High-content live-cell time-lapse imaging predicts cells about to die via apoptosis

Cell death is a dynamic process that unfolds through time. Live-cell time-lapse imaging captures these dynamics in a way thats impossible for static snapshots. High-content imaging (HCI), which has been developed for static microscopy, applied to time-lapse imaging can quantify how single-cell states change through time. Here we show the ability of high-content live-cell time-lapse imaging (HCLTI) to quantify the onset and progression of one form of cell death called apoptosis. We apply the Live Cell Painting assay called ChromaLIVETM and develop an HCLTI analysis pipeline. We show that HCLTI can discern the morphology dynamics of cells undergoing apoptosis, and demonstrate that machine learning can predict apoptosis as early as 100 minutes after exposing HeLa cells to the apoptosis inducer Staurosporine. This technical advancement paves the way for future studies to better understand the dynamics of other forms of cell death. Understanding cell death dynamics is one piece of solving larger biomedical puzzles like understanding how cells resist death (e.g., therapeutic resistance of cancer cells) and how cells die too soon (e.g., neurodegeneration).

systems biology↗

bioRxiv2018

Reevaluation of the Role of ERK3 in Perinatal Survival and Post-Natal Growth Using New Genetically-Engineered Mouse Models

The physiological functions of the atypical MAP kinase ERK3 remain poorly characterized. Previous analysis of mice with a targeted insertion of the lacZ reporter in the Mapk6 locus (Mapk6lacZ) showed that inactivation of ERK3 in Mapk6lacZ mice leads to perinatal lethality associated with intrauterine growth restriction, defective lung maturation, and neuromuscular anomalies. To further explore the role of ERK3 in physiology and disease, we generated novel mouse models expressing a catalytically-inactive (Mapk6KD) or conditional (Mapk6{Delta}) allele of ERK3. Surprisingly, we found that mice devoid of ERK3 kinase activity or expression survive the perinatal period without any observable lung or neuromuscular phenotype. ERK3 mutant mice reached adulthood, were fertile and showed no apparent health problem. However, analysis of growth curves revealed that ERK3 kinase activity is ncessary for optimal post-natal growth. To gain insight into the genetic basis underlying the discrepancy in phenotypes of different Mapk6 mutant mouse models, we analyzed the regulation of genes flanking the Mapk6 locus by quantitative PCR. We found that expression of several Mapk6 neighboring genes is deregulated in Mapk6lacZ mice, but not in Mapk6KD or Mapk6{Delta} mutant mice. Our genetic analysis suggests that off-target effects of the targeting construct on local gene expression are likely to be responsible for the perinatal lethality phenotype of Mapk6lacZ mutant mice.

developmental biology↗