Figure 7.
PC4 exhibits distinct modifications in HCC, and PC4 loss sensitizes HCC cells to CDK4/6 inhibitors. (A) Western blot showing the indicated protein expressions between six paired human HCC and adjacent tissue samples. N, normal. T, tumor. (B) Cell viability analysis by CCK8 assay in Huh7 and HepG2 cells treated with Palbociclib, siPC4, or a combination of Palbociclib and siPC4. Data are generated from n = 5 biological replicates. (C) Western blot shows the indicated protein expressions in Huh7 and HepG2 cells with Palbociclib, siPC4, or a combination of Palbociclib and siPC4. (D) Cell viability analysis by CCK8 assay in PC4-knockdown Huh7 and HepG2 cells treated with a combination of Palbociclib and PC4WT, PC4K68R, or PC4S17E. Data are generated from n = 5 biological replicates. (E) Western blot showing the indicated protein expressions in PC4-knockdown Huh7 and HepG2 cells treated with a combination of Palbociclib and PC4WT, PC4K68R, or PC4S17E. (F) Schema for Palbociclib, CLANsiPC4, or combination of Palbociclib and CLANsiPC4-treated Huh7-Luc xenografts model. (G) Bioluminescence from Huh7-Luc xenografts mice treated in F. (H) Photograph of tumors from Huh7-Luc xenografts mice treated in F. (I) Quantification of the tumor volume in Huh7-Luc xenografts mice treated in F. Data were generated from n = 6 in each group. (J) Overall survival from Huh7-Luc xenografts mice treated in F. Data are generated from n = 9 in each group. (K) Bioluminescence (top), photographs (middle), and H&E staining (bottom) of lungs from CLANNC, Palbociclib, CLANsiPC4, or Palbociclib+CLANsiPC4-treated Huh7-Luc xenografts mice. Scale bar = 1 mm. (L) Quantification of metastatic lesions to lung in K. Data are generated from n = 6 in each group. (M) Tunnel and Ki67 staining from Huh7-Luc xenografts mice treated in F. Data are generated from n = 6 in each group. Scale bar = 75 μm. (N) Representative images of CCND1, p-CDK2, p-RB, and CCND1 immunostaining from Huh7-Luc xenografts mice treated in F. Scale bar = 20 μm. Data are generated from n = 6 in each group. All quantifications are shown as means ± SD (one-way ANOVA test); error bars represent SD. **P < 0.01, ***P < 0.001, ****P < 0.0001. A, C, and E are representative of three independent experiments. Source data are available for this figure: SourceData F7.

PC4 exhibits distinct modifications in HCC, and PC4 loss sensitizes HCC cells to CDK4/6 inhibitors. (A) Western blot showing the indicated protein expressions between six paired human HCC and adjacent tissue samples. N, normal. T, tumor. (B) Cell viability analysis by CCK8 assay in Huh7 and HepG2 cells treated with Palbociclib, siPC4, or a combination of Palbociclib and siPC4. Data are generated from n = 5 biological replicates. (C) Western blot shows the indicated protein expressions in Huh7 and HepG2 cells with Palbociclib, siPC4, or a combination of Palbociclib and siPC4. (D) Cell viability analysis by CCK8 assay in PC4-knockdown Huh7 and HepG2 cells treated with a combination of Palbociclib and PC4WT, PC4K68R, or PC4S17E. Data are generated from n = 5 biological replicates. (E) Western blot showing the indicated protein expressions in PC4-knockdown Huh7 and HepG2 cells treated with a combination of Palbociclib and PC4WT, PC4K68R, or PC4S17E. (F) Schema for Palbociclib, CLANsiPC4, or combination of Palbociclib and CLANsiPC4-treated Huh7-Luc xenografts model. (G) Bioluminescence from Huh7-Luc xenografts mice treated in F. (H) Photograph of tumors from Huh7-Luc xenografts mice treated in F. (I) Quantification of the tumor volume in Huh7-Luc xenografts mice treated in F. Data were generated from n = 6 in each group. (J) Overall survival from Huh7-Luc xenografts mice treated in F. Data are generated from n = 9 in each group. (K) Bioluminescence (top), photographs (middle), and H&E staining (bottom) of lungs from CLANNC, Palbociclib, CLANsiPC4, or Palbociclib+CLANsiPC4-treated Huh7-Luc xenografts mice. Scale bar = 1 mm. (L) Quantification of metastatic lesions to lung in K. Data are generated from n = 6 in each group. (M) Tunnel and Ki67 staining from Huh7-Luc xenografts mice treated in F. Data are generated from n = 6 in each group. Scale bar = 75 μm. (N) Representative images of CCND1, p-CDK2, p-RB, and CCND1 immunostaining from Huh7-Luc xenografts mice treated in F. Scale bar = 20 μm. Data are generated from n = 6 in each group. All quantifications are shown as means ± SD (one-way ANOVA test); error bars represent SD. **P < 0.01, ***P < 0.001, ****P < 0.0001. A, C, and E are representative of three independent experiments. Source data are available for this figure: SourceData F7.

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