Fig. 3: Dissecting the molecular mechanisms underlying PELO dependency in 9p21.3−/− and MSI-H cancers.
From: SKI complex loss renders 9p21.3-deleted or MSI-H cancers dependent on PELO
a, Left, CRISPR knockout (KO) screen targeting 9p21.3 genes in KP4 cells with CRISPRi knockdown using Ch2-2 gRNA (control), or PELO gRNA 1 or 2. Right, differences in Chronos gene effect scores between PELO and Ch2-2 knockdown for both PELO gRNAs. b, Top, immunoblots of FOCAD, PELO and GAPDH in WM793 cells. Bottom, relative viability of WM793 cells with Ch2 gRNA or FOCAD knockout ± DOX-induced PELO knockdown. c, Top, immunoblots of FOCAD, PELO and α-tubulin in MIA PaCa-2 cells. Bottom, relative viability in MIA PaCa-2 cells stably expressing indicated cDNA ± DOX-induced PELO knockdown. d, Pearson correlation of microsatellite site length with PELO dependency scores in MSI-H cells (n = 73 cell lines). e, Cell lines (n = 1,100) plotted by PELO dependency and length of TTC37 intron 29 microsatellite repeats. f, Top, immunoblots of TTC37, PELO and β-actin in KP4 cells. Bottom, relative viability of KP4 ± TTC37 knockout ± DOX-induced PELO knockdown cells. g, Top, immunoblots of TTC37, PELO and β-actin in HCT116. Bottom, relative viability of HCT116 cells stably expressing the indicated cDNA ± DOX-induced PELO knockdown. Data are mean ± s.e.m. of biological replicates: n = 2 in a; n = 3 in b, f and g; and n = 3 except for PELO cDNA in DOX− condition, for which n = 2, in c. Significance was calculated as follows: in a, left-tailed Wilcoxon rank-sum test; in b and f, left-tailed Student’s t-test; in c and g, right-tailed Student’s t-test. Representative data from one experiment are shown. All experiments were performed twice, except for the experiment in a, which was performed once. For immunoblots, GAPDH, α-tubulin and β-actin were used as loading controls. Neg. ctrl, negative control; Pos. ctrl, positive control.
