Extended Data Fig. 3: Pharmacological or genetic inhibition of CDC7 induces a senescent phenotype in liver cancer cells with TP53 mutations.

a, b, Liver cancer cell lines with TP53 mutations (Hep3B, Huh7, SNU398, MHCC97H and HCCLM3) (blue) and liver cancer cell lines with wild-type TP53 (SK-Hep1 and Huh6) (red) were seeded at low confluence and grown in the absence or presence of the CDC7 inhibitors LY3177833 or TAK-931 at the indicated concentrations, in long-term colony-formation assays. Cells were fixed, stained and photographed after 10–14 days of culture. c, d, Growth curves (measured by Incucyte live-cell analyses) of liver cancer cell lines with TP53 mutations (Hep3B and Huh7) (blue) and liver cancer cell lines with wild-type TP53 (SK-Hep1 and Huh6) (red) exposed to LY3177833 or TAK-931. Graphs represent mean ± s.d. from four technical replicates. e, f, Liver cancer cells were cultured in the presence of the CDC7 inhibitors LY3177833 or TAK-931 at the indicated concentration for 4 days. SA-β-gal staining revealed that CDC7 inhibitors (LY3177833 or TAK-931) selectively induced senescence in liver cancer cells with TP53 mutations (blue) and not in liver cancer cells with wild-type TP53 (red). g, Liver cancer cell lines with TP53 mutations (Hep3B and Huh7) and liver cancer cell lines with wild-type TP53 (SK-Hep1 and Huh6) were stably transduced with control pLKO vector or with two independent shRNAs that target CDC7 (labelled here shCDC7 #1 and #2) and the efficiency of CDC7 knockdown in liver cancer cell lines was evaluated by western blot. h, Colony-formation assays of liver cancer cell lines with TP53 mutation (blue) and liver cancer-cell lines with wild-type TP53 (red), with and without CDC7 knockdown, were performed. Cells were fixed, stained and photographed after ten days of culture. i, CDC7 knockdown induced senescence in Hep3B and Huh7 cells with TP53 mutations, but not in SK-Hep1 and Huh6 cells, which have wild-type TP53. Senescence was detected by SA-β-gal staining. For gel source images, see Supplementary Fig. 1. Data in a–i are representative of three independent biological experiments.