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. 2024 Feb 13;223(3):e202308066. doi: 10.1083/jcb.202308066

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© 2024 Pan et al.

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Figure S6. — PC4 promotes G1–S transition and cell proliferation partially through a CCND1-dependent manner. Relative to Fig. 5. (A) Western blot showing PC4 and CCND1 protein expressions in Huh7 cells with control (NC), PC4 knockdown (siPC4), CCND1 knockdown (siCCND1), or combination knockdown of PC4 and CCND1 (siPC4+siCCND1). (B) Western blot showing CCND1 protein expression in HepG2 cells with control (NC) or CCND1 knockdown (siCCND1). (C) Cell viability analysis by CCK8 assay in HepG2 cells with NC or siCCND1. Data are generated from n = 5 biological replicates. (D) Western blot showing indicated protein expressions in Huh7 and HepG2 cells with NC, shPC4, or shPC4+CCND1. (E) Representative data and quantification of colony formation in colony assay in Huh7 and HepG2 cells transfected with NC, shPC4, or shPC4+CCND1. Data are generated from n = 3 biological replicates. (F) Representative data and quantification of soft agar colonies formation in soft agar assay in Huh7 and HepG2 cells transfected with NC, shPC4, or shPC4+CCND1. Scale bar = 300 μm. Data are generated from n = 4 biological replicates. (G) Representative data and quantification of EdU-positive cells in EdU assay in Huh7 and HepG2 cells transfected with NC, shPC4, or shPC4+CCND1. Scale bar = 75 μm. Data are generated from n = 3 biological replicates. (H) Western blot showing indicated protein expressions in Huh7 and HepG2 cells with NC, shPC4, or shPC4+SKP2. (I) Cell viability analysis by CCK8 assay in Huh7 cells with NC, shPC4, and shPC4+SKP2. Data were generated from n = 5 biological replicates. All quantifications were shown as means ± SD (one-way ANOVA test); error bars represent SD. **P < 0.01, ***P < 0.001, ****P < 0.0001. A, B, D, and H are representative of three independent experiments. Source data are available for this figure: SourceData FS6.