Fig 2: Induce and detect HCC827 and HCC827-AR cells. (A) HCC827 and HCC827-AR cells of cell viability assays are shown. (B) Protein expression levels of BIRC5 in HCC827 and HCC827-AR cells. (C) Effects of afatinib and YM155 on HCC827-AR cells as observed via cell viability assays are shown. ***p < 0.001.

Fig 3: Genes co-expressed with BIRC5 in LUAD (LinkedOmics). (A) The genes positively and negatively correlated with BIRC5 in LUAD. Heat maps showing top 50 genes positively (B) and negatively (C) correlated with BIRC5 in LUAD. Red indicates positively correlated genes and blue indicates negatively correlated genes. (D) Significantly enriched GO annotations and KEGG pathways of BIRC5 in TCGA cohort (E). The correlation between the hub genes and BIRC5 in LUAD (F).

Fig 4: Knockdown BIRC5. BIRC5 siRNA resulted in a partial decrease of cell viability (402-91, 69%; 1765-92, 55%; and DL-221, 69%). The transfection was successful as all three MLS cell lines and the control siRNA GAPDH showed a minor reduction of cell viability, whereas transfection with the positive control siPLK1 resulted in all three cell lines with a pronounced decrease in cell viability. The transfection reagent alone hardly affected cell viability (A). A reduction in GAPDH and survivin protein expression was found after transfection with, respectively, siGAPDH and siBIRC5 in all three cell lines, although knockdown efficiency was variable (B).

Fig 5: Trp53R172H, not wild-type p53, binds to YAP and fosters Survivin expression in murine ESCC cells. (A, left) Endogenous expression level of YAP and p53 in wild type, Trp53R172H/− shCtrl, shTrp53, Trp53−/− Ctrl, and Trp53−/− R172H cells. (Right) Expression levels of YAP and p53 in LS123 nontarget control (NT), Crispr knockout (Crispr) cell lines, and H1299 empty vector control (control) and p53R175H (R175H) ectopic expressed cell lines. (B) Proximity ligation assay (PLA) for YAP and Trp53R172H. Scale bar, 50 µm. (C) Quantification of the average number of PLA signals shows significantly high levels of signals in Trp53R172H/− shCtrl cells (vs. shTrp53 cells, n > 100 cells per group, P < 0.0001, unpaired t-test, error bars represent SEM) and Trp53−/− R172H cells (vs. Ctrl cells, n > 100 cells per group, P < 0.0001, unpaired t-test, error bars represent SEM). (D) Immunoprecipitation-Western blot analysis of Trp53R172H-HA with YAP-DDK in 293 FT cells. (E) Chromatin immunoprecipitation (ChIP) with YAP antibody in WT1, Trp53R172H/− shCtrl, and shTrp53 cells demonstrates increased level of YAP on the BIRC5 promoter in the presence of Trp53R172H. shCtrl versus WT1 cells, n = 3 replicates per group, P = 0.0052, unpaired t-test; shCtrl versus shTrp53 cells, n = 3 replicates per group, P = 0.0107, unpaired t-test. Error bars represent SEM. (F) Proximity ligation assay for YAP and p53R175H. Scale bar, 20 µm. (G) Quantification of the average number of PLA signals shows significantly high levels of signals in LS123 NT cells (vs. Crispr cells, n > 100 cells per group, P < 0.0001, unpaired t-test, error bars represent SEM) and H1299 R175H cells (vs. control cells, n > 100 cells per group, P < 0.0001, unpaired t-test, error bars represent SEM).