Fig. 7
CDC16 overexpression rescued cell cycle arrest in G1 stage induced by c-Jun overexpression. (A-B). Cell cycle analysis by flow cytometry for negative control (NC), c-Jun OE and c-Jun OE + CDC16 OE cells. Percentage of cells at different cell cycle stage was analyzed with ordinary one-way ANOVA combined with Sidak’s multiple comparisons (mean ± SEM, n = 3; *p < 0.05, **p < 0.01). (C). Schematic representation of cell cycle progression and expression of cyclins at different stage. c-Jun overexpression inhibited cell cycle progression from G1 to S stage, increased G1 to G0 stage and promoted cell differentiation, whereas c-Jun and CDC16 co-overexpression promoted cell cycle progression. (D). Western blot showing the expression of cell cycle related markers in NC, c-Jun OE and C-Jun OE + CDC16 OE cells. Relative protein level was analyzed as ratio to GAPDH and heatmap was represented as mean ± SEM (n = 3). (E). qPCR result showing the mRNA expression of cell cycle related markers (cyclin D1, cyclin E1, cyclin A2, cyclin B and CDKs) in NC, c-Jun OE and c-Jun OE + CDC16 OE cells. Data were analyzed with ordinary one-way ANOVA combined with Sidak’s multiple comparisons (mean ± SD, n = 3; *p < 0.05, **p < 0.01, ***p < 0.001). (F). CDC16 shRNA was constructed to pLVX-shRNA2-GFP plasmid and then transfect into HEK293T cells. Cells expressing pLVX-shRNA2-GFP-CDC16 shRNA showed green fluorescence and expression of CDC16 was detected by Western blot. (G). Represent images of SH-SY5Y cells transfected with CDC16 shRNA lentivirus (left: 100µL and right: 200µL). (H-I). Western blot showing the expression of cell cycle related markers in NC, c-Jun OE and c-Jun OE + CDC16 knockdown cells. Relative protein level was analyzed as ratio to GAPDH (mean ± SEM, n = 3; *p < 0.05, **p < 0.01, ***p < 0.001)