Fig 2: XIAP-RIPK2-TAB1 axis supports neutrophil infiltration and melanoma growth in B16 mouse melanoma Intensities of CD177 and XIAP immunostainings in tumour sections from 55 patients were visually scored. The numbers of melanomas with different intensities were plotted into the table as a percentage of the total. Specific staining intensity for XIAP and CD177, corresponding to relative amounts of infiltrated neutrophils, was arbitrarily set as the following: -, not expressed; +, low; ++, moderate; and +++, strong expression. Representative IHC pictures (right panel). Scale bar 100 µm.Representative IF (neutrophils staining) in B16F1 WT, XIAPKO, TAB1KO or RIPK2KO tumours (Scale bar 50 µm) and quantification of Ly6G+ cells. The sum of Ly6G-positive cells from 5 randomly selected areas of the tumour (5 mice per genotype) was represented. Dots represent individual mice (n = 5).B16F1 WT, XIAPKO (clone 2) subcutaneous melanoma tumour growth in BL/6 WT mice (n = 5). Mice were intra-peritoneal injected with IgG antibody as a negative control or Ly6G antibody 1 day prior to the subcutaneous injection of the B16F1 WT or XIAPKO cells. Data are mean ± s.e.m. Two-way analysis of variance (ANOVA) followed by Dunnett’s post-analysis.Representative IF (neutrophils) scale bar 50 µm and quantification of Ly6G+ cells in tumours from C. The sum of Ly6G-positive cells from 5 randomly selected areas of the tumour (4 mice per genotype) was represented. Dots represent individual mice (n = 4). Data information: In B and D, data are presented as mean ± s.e.m. One-way analysis of variance (ANOVA) followed by Dunnett’s post-analysis. **P < 0.01; ***P < 0.001; ****P < 0.0001 and ns, not significant.

Fig 3: WSTF-RPB1 is strongly associated with the anti-AML activity. Triptolide downregulates the WSTF and RPB1 signaling pathway without affecting TAB1/P38 signaling in a dose dependent manner in MV-4-11 (A) and THP-1 (B). Time-dependent experiments show that triptolide inhibits the WSTF-RPB1 signaling pathway with no effect on TAB1/P38 signaling in MV-4-11 (C) and THP-1 (D). (E) The different expression levels of WSTF, XPB, and RPB1 is shown.

Fig 4: XIAP BIR1- and -BIR2 support melanoma growth via TAB1- and RIPK2-dependent signalling B16F1 WT, B16F1-XIAP?BIR1 and B16F1-XIAP?BIR2 subcutaneous melanoma tumour growth in BL/6 WT mice (n = 3 each genotype).B16F1 WT, B16F1-XIAPKO (clone 1 and 2), B16F1-TAB1KO and B16F1-RIPK2KO (clone 1 and 2) subcutaneous melanoma tumour growth in BL/6 WT mice (n = 5 each genotype).qRT–PCR analysis of different cytokines/chemokines in the indicated melanoma tumours in mice. Dots represent individual mice (n = 3). Data are presented as mean ± SD. One-way analysis of variance (ANOVA) followed by Dunnett’s post-analysis. Data information: In A and B data are mean ± s.e.m; two-way analysis of variance (ANOVA) followed by Dunnett’s post-analysis. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 and ns, not significant.

Fig 5: Thrombin Activation of p38 MAPK Requires c-Src:(A and C) Immunoblot of cell lysates from EA.hy926 cells pretreated with Ctrl, PP2, or PP3 (A) or Ctrl and PP2 (C), prior to a-Th stimulation. The data (mean ± SD, n = 3) were analyzed by ANOVA (*p < 0.05, **p < 0.01, ***p < 0.001).(B and D) Immunoblots of EA.hy926 cell lysates transfected with ns- or c-Src-specific siRNA prior to a-Th stimulation immunoblotted for p38 and ERK1/2 (B) or TAB1 (D). The data (mean ± SD, n = 3) were analyzed by ANOVA (*p < 0.05, **p < 0.01).