Fig. 4: Scaf1 functions as PDAC tumor suppressor.
From: In vivo CRISPR screens reveal SCAF1 and USP15 as drivers of pancreatic cancer
A Tumor-free survival of Pdx1-Cre;LSL-KrasG12D;LSL-Cas9-GFP mice injected with CRISPR AAV targeting the indicated gene or non-targeting control sgRNA (sgCtrl n = 6). Two independent sgRNAs were used (sgScaf1_1 n = 8, sgScaf1_2 n = 7). Log-Rank test (Mantel-Cox). B Representative H&E images showing multifocal PanINs in sgCtrl-transduced pancreas and PADC tumors in sgScaf1-transduced pancreas. Scale bar 100 µm. C Cell proliferation curves of KC sgCtrl and sgScaf1 cells were obtained using the IncuCyte live-cell imaging and data are expressed as cell confluence percentage (%; mean ± SD, n = 3 independent experiments). Two-way ANOVA (sgScaf1_1 p = 0.0039: sgScaf1_2 p = 0.00042), Dunnett’s multiple comparison. D Tumor-free survival of NSG mice orthotopically injected with sgCtrl (n = 5) or sgScaf1_1/2 (n = 5; n = 5) KC cells. Two independent sgRNAs were used. Log-Rank test (Mantel-Cox). E Dose-response curves for KPC sgCtrl or sgScaf1 cells treated with the indicated concentration of Olaparib (mean ± SD, n = 3 independent experiments). Two-way ANOVA (sgScaf1_1 p = 0.0084: sgScaf1_2 p = 0.0028), Dunnett’s multiple comparisons. F Representative Western Blot of Usp15 in KC cells transduced with the indicated sgRNAs and treated as indicated. This experiment was repeated independently two times with similar results. G Cell growth curves of KC sgCtrl and sgScaf1 cells expressing the listed isoform of USP15 or an empty vector (EV). Data are expressed as cell confluence percentage (%; mean ± SD, n = 3 independent experiments); two-way ANOVA (sgScaf1+EV p = 0.0342), Dunnett’s multiple comparison. Dose-response curves for KC sgCtrl and sgScaf1 cells expressing the listed isoforms of USP15 or EV and treated with Olaparib. (%; mean ± SD, n = 3 independent experiments; two-way ANOVA (sgScaf1+EV p = 0.00129), Sidak multiple comparison H Tumor-free survival of NSG mice orthotopically injected with sgCtrl (n = 5) or sgScaf1 KC cells expressing the listed isoforms of USP15 (n = 5: n = 5) or EV (n = 5; n = 5). Log-Rank test (Mantel-Cox).
