Fig. 4: Cellular health and cell-painting phenotypes are correlated.
From: Reference compounds for characterizing cellular injury in high-content cellular morphology assays
Compounds tested in cell painting (CP) were profiled for cellular health biomarkers under CP-like conditions to better understand the relationship between CP phenotypes and cellular health. a CP activities and relative cell numbers correlate with cellular confluence, caspase 3/7 activation, cell viability, and overlap (cells with caspase 3/7 activation and loss of membrane integrity) after 24-h compound treatment as measured by live-cell imaging. AUCs were calculated from the live-cell imaging assay concentration-response curves for each compound. b Breakdown of live-cell imaging cellular health profiles (AUC) by CP phenotypic clusters and cell injury compound groups. The CP phenotypic clusters associated with prototypical cell injury compounds have decreased cellular confluence and increased caspase 3/7 activation and loss of membrane integrity. The cellular health biomarkers vary depending on compound group. Object count indicates the number of cells (“objects”) above the signal threshold for each biomarker. The top/bottom of the boxes refer to the first/third quartiles; the whiskers refer to either the most extreme value or 1.5× inter-quartile range from the first/third quartiles, whichever has a smaller absolute value. c Select live-cell imaging profiles for cellular health. Cell injury compounds predictably decrease confluence, and can lead to increases in caspase 3/7 activation and membrane disruption within 24 h of compound treatment. Data are mean ± SD of three intra-run technical replicates each performed on separate microplates. d Cell injury compounds, most notably electrophiles, decrease the intracellular glutathione (GSH):oxidized glutathione (GSSG) ratio. Black, not tested; yellow, outlier (GSSG > 200%) or not calculated (GSH:GSSG, calculated GSH < 0%). Data are mean (±SD) of five intra-run technical replicates each performed on separate microplates. h/ngKATI; historical/next-generation lysine acetyltransferase inhibitor; NSE-(IA), nonspecific electrophiles (inactive analog); TE, targeted electrophile. Source data are provided as a Source Data file.
