Fig. 7: Influence of YAP and β-catenin on OIS.
From: Alternative Wnt-signaling axis leads to a break of oncogene-induced senescence
A Percentages of SA-β-Galactosidase positive cells (blue) in SBcl2, WM1366, and MV3 cell lines 48 h after transfection with siYAP or siCtr (left) (n = 3). Bars are shown as mean ± SEM (multiple T-tests (Bonferroni-Dunn) *P < 0.05, ns: not significant). Sample image of light microscopic examination of SA-β-Galactosidase staining in SBcl2 cells (right). B Percentages of SA-β-Galactosidase positive cells (blue) in SBcl2, WM1366, and MV3 cell lines 48 h after transfection with siβ-catenin 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). CImmunofluorescence staining of PML and DAPI in melanoma cell lines 48 h after the transfection with siYAP or siCtr. The graph shows the number of nuclear PML bodies (left). Representative image of overlays of PML (red) and DAPI (blue) in MV3 cells (right). D Immunofluorescence staining of PML and DAPI in melanoma cell lines 48 h after the transfection with siβ-catenin 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 MV3 cells (right). E Cell cycle analysis 48 h after YAP 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 MV3 cells 48 h after siYAP or siCtr transfection. F Cell cycle analysis 48 h after β-catenin 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 MV3 cells 48 h after siβ-catenin or siCtr transfection. G Percentages of SA-β-Galactosidase positive cells (blue) in mock/BRAFm-, mcherry/mcherry β-cateninS33Y OE double-transduced NHEMs (left) (n = 3). Bars are shown as mean ± SEM (two-way ANOVA (Tukey) *P < 0.05, ns: not significant). Sample image of light microscopic examination of SA-β-Galactosidase staining (right). H Immunofluorescence stainings of PML and DAPI in mock/BRAFm-transduced, mcherry/mcherry β-cateninS33Y OE double transduced NHEMs. The graph shows nuclear accumulation of PML (n = 3). Bars are shown as mean ± SEM (two-way ANOVA (Tukey) *P < 0.05, ns: not significant). I Representative real-time cell proliferation curves of BRAFm, mcherry/mcherry β-cateninS33Y OE double transduced NHEMs (left panel) and quantified “slope” (proliferative ability) (right panel) (n = 3). Bars are shown as mean ± SEM (two-way ANOVA (Tukey) *P < 0.05, ns: not significant).
