Fig. 4
PEL cell lines are dependent on CCND2 and CFLAR. a Heatmap of adj. p values of sgRNA depletion of CCND2 and CFLAR in cell lines screened. Indicated on the right are the numbers of cancer types (out of 16) where the gene was found to be a potential dependency. The Brunello library was used for most of the screens except where indicated: G, GeCKO v2. b, c Knockout of CFLAR. b Representative analysis of relative live BCBL-1 cell numbers over time following CFLAR knockout. n = 4. c Representative western blots of c-FLIPL and c-FLIPS isoforms for b. d, e Similar to b and c but using CCND2 sgRNAs. Experiments in b–e were performed together and thus share controls. f Distribution of cell cycle phase populations in BCBL-1 Cas9 cells upon CCND2 or CFLAR knockout analyzed by propidium iodide staining of samples on day 4 in experiments shown in b–e. p Values were calculated by Student’s t test and compared to sgAAVS1. g Calculated IC50 values of palbociclib in the indicated cell lines. Gray bars, non-PEL cells; pink bars, PEL cell lines; n ≥ 3. h, i Cell cycle analysis of propidium iodide-stained live BCBL-1 cells treated for 24 h with 220 nM palbociclib (IC50). h Representative histograms of DNA content from propidium iodide staining. i Distribution of cell cycle phase populations. p Values were calculated by Student’s t test and compared to PBS-treated cells. n = 3. All error bars, SEM
