Fig. 4: Mitochondrial priming and response to BH3 mimetics in BAX-incompetent TIS models of LoVo colon cancer cells.
From: Mitochondrial priming and response to BH3 mimetics in “one-two punch” senogenic-senolytic strategies
A Left. BH3 profiling was performed to measure mitochondrial depolarization in proliferative (untreated) and TIS LoVo colon cancer cells exposed to activator and sensitizer BH3 peptides. Figure shows heat maps of % mitochondrial depolarization caused by increasing concentrations of the activator (BIM, BID, PUMA) and sensitizer (BMF, BAD, NOXA, HRK) peptides in proliferative (untreated) and palbociclib and bleomycin TIS cells. Data shown are the mean of ≥3 independent experiments using three technical replicates for each peptide. Graphs represent the means (columns) ± S.E.M. (bars) of ≥3 independent experiments BH3 peptide EC50s (μmol/L) in proliferative (untreated) and palbociclib and alisertib TIS cells. Statistically significant differences (ANOVA analysis) between proliferative and TIS phenotypes means are shown. n.s. not statistically significant. Right. Figure shows global heat maps of % mitochondrial depolarization caused by selected concentrations of activator and sensitizer peptides in LoVo colon cancer cells. Samples are ordered according to depolarization by the BIM peptide. Data shown are the mean of ≥3 independent experiments with three technical replicates for each peptide. B TIS cancer cells and proliferative (untreated) cells were replated into 96-well plates (7,000 cells/well and 4,000 cells/well, respectively) and cultured with increasing concentrations of ABT-263/navitoclax, A1331852, ABT-199/venetoclax, or S63845. After 5 days, cell viability was measured using the alarmarBlue™ assay. Shown are representative images of alamarBlue™-based cell viability assays showing changes in the number of metabolically active cells in response to serial dilutions of BH3 mimetics and dose-response curves for ABT-263/navitoclax, A1331852, ABT-199/venetoclax, or S63845 in proliferative (untreated) and TIS LoVo cancer cells (untreated=100%). See also Table 2. C Top. LoVo colon cancer cells were treated with senescence-inducing concentrations of alisertib or palbociclib for 7 days. Senescent cells and parental (proliferative) cells were replated into 12-well plates (200,000 cells/well and 150,000 cells/well, respectively) and cultured with increasing concentrations of ABT-263/navitoclax, A1331852, ABT-199/venetoclax, or S63845. After 5 days, cell growth and senescence were monitored using crystal violet staining (left panels) and SA-β-gal staining (right panels). Shown are representative images from three technical replicates. Scale bar = 200 μm. Bottom. Summary of the responses of LoVo TIS cancer cells to BH3 mimetics.
