Fig. 4: Role of Wnt10b on OIS and in melanoma cells.
From: Alternative Wnt-signaling axis leads to a break of oncogene-induced senescence
A Representative immunohistochemical staining of Wnt10b protein in normal human skin, primary human melanoma, and human metastatic melanoma tissue samples (n = 10). B Percentages of SA-β-Galactosidase positive cells (blue) in mock/BRAFm-transduced NHEMs treated with recombinant Wnt10b or PBS (left) (n = 3). Bars are shown as mean ± SEM (two-way ANOVA (Tukey) *P < 0.05, ns: not significant). Example image of light microscopic examination of SA-β-Galactosidase staining (right). C Immunofluorescence staining’s of PML and DAPI in mock/BRAFm-transduced NHEMs treated with recombinant Wnt10b or PBS. The graph shows nuclear accumulation of PML (left) (n = 3). Bars are shown as mean ± SEM (two-way ANOVA (Tukey) *P < 0.05, ns: not significant). Sample image of overlays of PML (red) and DAPI (blue) (right). D Representative real-time cell proliferation curves of BRAFm-transduced NHEMs treated with recombinant Wnt10b or PBS (BRAFm PBS set as 1) (right panel) and quantified “slope” (proliferative ability) (left panel) (n = 3). Bars are shown as mean ± SEM (multiple T-tests (Bonferroni-Dunn) *P < 0.05, ns: not significant). E Percentages of SA-β-Galactosidase positive cells (blue) in SBcl2, WM1366, and MV3 cell lines 48 h after transfection with siWnt10b or siCtr (left) (n = 3). Bars are shown as mean ± SEM (multiple T-tests (Bonferroni-Dunn) *P < 0.05, ns: not significant). Example image of light microscopic examination of SA-β-Galactosidase staining in SBcl2 cells (right). F Immunofluorescence staining of PML and DAPI in melanoma cell lines 48 h after the transfection with siWnt10b or siCtr. The graph shows the number of nuclear PML bodies (left) (n = 3). Bars are shown as mean ± SEM (multiple T-tests (Bonferroni-Dunn) *P < 0.05, ns: not significant). Sample image of overlays of PML (red) and DAPI (blue) in SBcl2 cells (right). G Cell cycle analysis 48 h after Wnt10b inhibition in SBcl2, WM1366, and MV3 cells. Bars represent cells in G1 phase (left) (n = 3). Bars are shown as mean ± SEM (multiple T-tests (Bonferroni-Dunn) *P < 0.05, ns: not significant). Representative histograms of cell cycle analysis in SBcl2 cells 48 h after siWnt10b or siCtr transfection.
