Fig 2: XIAP promotes tumour growth without interfering with tumour cell apoptosis ARepresentative Incucyte images and cell death measurements using Sytox Green in B16F1 WT and XIAPKO (clone 1 and 2) cells after 24 h treatment with 100 ng/ml TNF. DMSO was used as a negative control. Scale bar 200 µm. 2 mM H2O2 was used as a positive control. Scale bar 400 µm.BIncucyte cell death measurements using Sytox Green in B16F1 WT and XIAPKO (clone 1 and 2) cells after 24 h treatment with 50 ng/ml TRAIL. DMSO was used as a negative control. 2 mM H2O2 was used as a positive control.CCell proliferation analysis using CSFE intensity (FACS analysis) (left) or using Incucyte Live-Cell-Analysis assay (right) examining the proliferation of B16F1 WT and XIAPKO CRISPR–Cas9 clones.DqRT–PCR analysis of Cxcl1 (KC) and Cxcl10 in B16F1 WT and XIAPKO (clone 1 and 2) cells.EWestern blot analysis of cell lysates of BLM WT, XIAPKO, SK-Mel28 WT, SK-Mel28-XIAPKO, Colo38 WT, Colo38-XIAPKO, HCT116 WT and HCT116-XIAPKO after CRISPR–Cas9 gene editing.F, GIL8 measurement in the supernatant of SK-Mel28 and M5 after 48 h siRNA transfection (F) or after 24 h of seeding BLM, BLM-XIAPKO, SK-Mel28, and SK-Mel28-XIAPKO cells (G). Cells were treated with 20 ng/ml TNF for 8 h. Data information: In A, B and C (right panel) data are mean ± s.e.m; two-way analysis of variance (ANOVA) followed by Tukey’s post-analysis. Data derived from three individual biological replicates. In C (left), D, F and G, data are presented as mean ± SD. One-way analysis of variance (ANOVA) followed by Dunnett’s post-analysis. Dots represent individual biological replicates. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 and ns, not significant. Source data are available online for this figure.

Fig 3: The schematic diagram of this study: miR-137 suppresses XIAP expression to sensitize TRAIL-induced cell death in GBM.

Fig 4: miR-4463 promotes proapoptotic signaling in H2O2-treated HUVECs. (A) HUVECs were pretreated with miR-4463 mimics, a miR-4463 inhibitor or a NC miRNA, and then exposed to 700 µmol/l H2O2 for 16 h; Control cells did not receive treatment. (B) Western blot analysis was used to assess the protein expression levels of the proapoptotic factors PARP1, Bax and C-caspase3, and the antiapoptotic factors XIAP and Bcl-2 (n=3). *P<0.05 vs. 700+NC. Bax, Bcl-2-associated X protein; Bcl-2, B-cell lymphoma-2; C, cleaved; H2O2, hydrogen peroxide; HUVEC, human umbilical vein endothelial cell; miR, microRNA; NC, negative control; PARP1, poly (adenosine diphosphate-ribose) polymerase 1; XIAP, X-linked inhibitor of apoptosis protein.

Fig 5: Response to Birinapant treatments in colon cancer cell lines.A HCT116, LS1034, LS513, and MDST8 cells were treated for 24 h and 48 h with the indicated treatments. The colored bars represent the subtype of the cell lines (CMS1—orange; CMS2—blue; CMS3—pink; CMS4—green). Blots were probed for cIAP1, cIAP2, and XIAP as well as the cleaved forms of caspase-3, -7, and -8. B–D All cell lines were treated for 48 h with 10 ng/ml TNFa, 1 µM Birinapant, 2 µM oxaliplatin/ 10 µM 5-FU (Oxa/5-FU) or a combination of these drugs. B Gating strategy for apoptosis assay. Only the sum of necrotic, early-, and late-apoptotic cells are shown in C and D. C Combined treatment results of all cell lines by CMS subtype. To normalize cell line intrinsic cell death, the background cell death (DMSO control) was subtracted. A two-way ANOVA was performed, excluding the CMS4 subtype. A Tukey’s test for multiple comparisons was used for post hoc analysis. D Treatment outcomes per cell line (mean + SD), grouped by their subtypes. To highlight the effects of Birinapant only, the single treatments and their corresponding co-treatment with Birinapant are shown. N = 3; *P = 0.05; **P = 0.01; ***P = 0.001.