Fig. 3: Effect of Wnt6 on OIS and its role in melanoma cells.
From: Alternative Wnt-signaling axis leads to a break of oncogene-induced senescence
A Representative immunohistochemical staining of Wnt6 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 co-cultured with Wnt6 OE HEK (Wnt6 CC) or with CMV OE HEK (CMV CC) cells (left) (n = 3). Bars are shown as mean ± SEM (two-way ANOVA (Tukey) *P < 0.05, ns: not significant). Exemplary image of light microscopic examination of SA-β-Galactosidase staining (right). C Immunofluorescence staining of PML and DAPI in mock/BRAFm-transduced NHEMs co-cultured with Wnt6 OE HEK (Wnt6 CC) or with CMV OE HEK (CMV CC) cells (n = 3). Bars are shown as mean ± SEM (two-way ANOVA (Tukey) *P < 0.05, ns: not significant). D Immunofluorescence staining of KI-67 and DAPI in mock/BRAFm-transduced NHEMs co-cultured with Wnt6 OE HEK (Wnt6 CC) or with CMV OE HEK (CMV CC) cells. The graph shows the nuclear accumulation of KI-67 (left) (n = 3). Bars are shown as mean ± SEM (two-way ANOVA (Tukey) *P < 0.05, ns: not significant). Sample image of overlays of KI-67 (purple) and DAPI (blue) (right). E Percentages of SA-β-Galactosidase positive cells (blue) in SBcl2, WM1366 and MV3 cell lines 48 h after transfection with siWnt6 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 siWnt6 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). Example image of overlays of PML (red) and DAPI (blue) in SBcl2 (right). G Cell cycle analysis 48 h after Wnt6 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 siWnt6 or siCtr transfection.
