Fig. 2: Focused validation of PELO dependency in vitro and in vivo.
From: SKI complex loss renders 9p21.3-deleted or MSI-H cancers dependent on PELO
a, Viability effect of CRISPRi-mediated PELO knockdown (KD) normalized to average of negative controls (grey dashed line; empty vector and/or gRNA Ch2-2) and positive controls (orange dotted line; gRNA for SF3B1 and/or POLR2D) for 9p21.3+ or short deletion/MSS (n = 4), 9p21.3−/−/MSS (n = 3) and 9p21.3+/MSI-H cell lines (n = 3). b, Top, immunoblots of PELO and vinculin in MIA PaCa-2 cells transduced with the indicated gRNA ± PELO cDNA. Bottom, viability scores normalized to average viability of negative controls. c, Immunoblots of PELO and vinculin levels in the indicated patient-derived tumour organoid models. Relative viability following DOX-induced knockdown of the indicated gRNA, relative to DMSO control. d, Average tumour volume over time for nude mice with subcutaneous engraftment of MIA PaCa-2 cells following randomization to a standard diet (control) or DOX-containing diet to induce PELO knockdown. e, Kaplan–Meier survival plot for mice randomized to DOX-containing (n = 5) or standard (n = 6) diet. Data are mean ± s.e.m. of biological replicates: n = 6 for all cell lines except KP4 and GB1, for which n = 7, and IGROV-1, for which n = 3 in a, n = 3 in b, n = 2 for PANFR0127 and PANFR0071 and n = 3 for CCLF_CORE_0001 in c; n = 9 tumours for standard diet and n = 10 for DOX-containing diet in d. Significance was calculated as follows: left-tailed Student’s t-test (a), right-tailed Student’s t-test (b), pairwise log-rank test (e). Representative data are shown from two experiments in a and one experiment in b–e. Experiments were performed twice for a–c and once for d and e. For immunoblots, vinculin was used as loading control.
