1. Fig. 2. Resistance to S63845-induced apoptosis through loss of BAK or elevated BCL-XL.
Resistance to S63845-induced apoptosis through loss of BAK or elevated BCL-XL. (A) SK-BR-3 cells were infected with a genome-wide lentiviral sgRNA library and treated with S63845 (1 μM) or DMSO control, and then gDNA of surviving cells was isolated to identify the sgRNAs by NGS. The pooled analysis from six independent infections, displaying normalized values for S63845 or DMSO control, is shown. Solid red bar represents the regression line. The sgBAK1 hits are shown as blue dots. See table S5 for top up-regulated sgRNAs in the S63845-treated pools. (B) SK-BR-3 cells infected with CRISPR-Cas9 guides targeting proapoptotic proteins were treated with increasing concentrations of S63845 for 24 hours before assessment of viability using CellTiter-Glo. Ev, empty vector. (C) SK-BR-3 cells infected with CRISPR-Cas9 guides targeting BIM, BID, and PUMA were treated with increasing concentrations of S63845, as described above. (D) The MDA-MB-468 cell line was treated with increasing concentrations of S63845 in the presence of ABT-199, WEHI-539, or ABT-737 (500 nM) for 24 hours before assessment of viability using CellTiter-Glo. For (B) to (D), means ± SEM for three independent experiments are shown. (E) PDX tumor cells were cultured for 24 hours in mammosphere medium with ABT-199, ABT-737, WEHI-539, and S63845 (500 nM and 1 μM) or combination treatment with S63845 and other BH3 mimetics (both at 500 nM) before assessment of viability using CellTiter-Glo. Results are presented as percentages of untreated cells. Bars represent means ± SEM for at least five independent experiments per PDX. The tumor subtype for each PDX is shown in parentheses.
Delphine Merino et al., Sci Transl Med 2017;9:eaam7049
Published by AAAS