Figure 6.
Phosphorylation of PC4 at S17 or deubiquitylation at K68 prolongs the G1–S transition and suppresses cell proliferation. (A) Asynchronized cell cycle analysis in PC4-knockdown Huh7 cells stably expressing indicated Flag-tagged PC4 variants. The number in the upper right corner represents the percentage of cells in different cell cycle phases. Data are generated from n = 3 biological replicates. (B) Representative frames of FUCCI-expressing Huh7 transfected with siRNA and PC4-mutant plasmid via time-lapse imaging. Scale bar = 50 μm. (C) The time length of the G1 phase in FUCCI-expressing Huh7 transfected with siPC4 and PC4-mutant plasmid via time-lapse imaging. Data are generated from n = 30 cells in each group. (D) Cell viability analysis by CCK8 assay in PC4-knockdown Huh7 cells transfected with indicated Flag-tagged PC4 variants. Data are generated from n = 5 biological replicates. (E) Representative images and quantification of colony formation in colony assays in PC4-knockdown Huh7 cells transfected with indicated Flag-tagged PC4 variants. Data are generated from n = 3 biological replicates. (F) Representative images of soft agar colony formation in soft agar assays and EdU-positive cells in EdU assays in PC4-knockdown Huh7 cells transfected with indicated Flag-tagged PC4 variants. Soft agar scale bar = 300 μm, EdU scale bar = 75 μm. (G) Quantification of soft agar colony formation in soft agar assays in PC4-knockdown Huh7 cells transfected with indicated Flag-tagged PC4 variants. Data are generated from n = 4 biological replicates. (H) Quantification of EdU-positive cells in EdU assays in PC4-knockdown Huh7 cells transfected with indicated Flag-tagged PC4 variants. Data are generated from n = 3 biological replicates. (I) Tumor volume in the nude mice subcutaneously injected with PC4-knockdown Huh7 cells stably expressing indicated Flag-tagged PC4 mutants. Data are generated from n = 3 biological replicates. (J) Representative photographs of dissected tumors and tumor weight in the nude mice subcutaneously injected with PC4-knockdown Huh7 cell stably expressing indicated Flag-tagged PC4 mutants. Data are generated from n = 5 biological replicates. All quantifications are shown as means ± SD (one-way ANOVA test); error bars represent SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Phosphorylation of PC4 at S17 or deubiquitylation at K68 prolongs the G1–S transition and suppresses cell proliferation. (A) Asynchronized cell cycle analysis in PC4-knockdown Huh7 cells stably expressing indicated Flag-tagged PC4 variants. The number in the upper right corner represents the percentage of cells in different cell cycle phases. Data are generated from n = 3 biological replicates. (B) Representative frames of FUCCI-expressing Huh7 transfected with siRNA and PC4-mutant plasmid via time-lapse imaging. Scale bar = 50 μm. (C) The time length of the G1 phase in FUCCI-expressing Huh7 transfected with siPC4 and PC4-mutant plasmid via time-lapse imaging. Data are generated from n = 30 cells in each group. (D) Cell viability analysis by CCK8 assay in PC4-knockdown Huh7 cells transfected with indicated Flag-tagged PC4 variants. Data are generated from n = 5 biological replicates. (E) Representative images and quantification of colony formation in colony assays in PC4-knockdown Huh7 cells transfected with indicated Flag-tagged PC4 variants. Data are generated from n = 3 biological replicates. (F) Representative images of soft agar colony formation in soft agar assays and EdU-positive cells in EdU assays in PC4-knockdown Huh7 cells transfected with indicated Flag-tagged PC4 variants. Soft agar scale bar = 300 μm, EdU scale bar = 75 μm. (G) Quantification of soft agar colony formation in soft agar assays in PC4-knockdown Huh7 cells transfected with indicated Flag-tagged PC4 variants. Data are generated from n = 4 biological replicates. (H) Quantification of EdU-positive cells in EdU assays in PC4-knockdown Huh7 cells transfected with indicated Flag-tagged PC4 variants. Data are generated from n = 3 biological replicates. (I) Tumor volume in the nude mice subcutaneously injected with PC4-knockdown Huh7 cells stably expressing indicated Flag-tagged PC4 mutants. Data are generated from n = 3 biological replicates. (J) Representative photographs of dissected tumors and tumor weight in the nude mice subcutaneously injected with PC4-knockdown Huh7 cell stably expressing indicated Flag-tagged PC4 mutants. Data are generated from n = 5 biological replicates. All quantifications are shown as means ± SD (one-way ANOVA test); error bars represent SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal