Fig. 2: PUM1 and PUM2 drive colorectal cancer cell growth in vitro and tumorigenicity in vivo.

a The relative levels of Pum1 and Pum2 mRNA in six human colorectal cancer cell lines normalized over their levels in a human colon epithelial cell line, NCM460. Actin was used as an internal control to normalize the level of Pum1 or Pum2 among the duplicate samples (n = 3) of the same cell line. Error bars represent SEM. n.s not significant, *P < 0.05, **P < 0.01, ***P < 0.001 by Student’s t-test. b Levels of PUM1 or PUM2 proteins in a normal colon epithelial cell line NCM460 and six human CRC cell lines. β-actin was used as a loading control. c Knockout efficiency of Pum1 and Pum2 in HCT116 and RKO cells, in which HCT116 Pum1^−/−-1 represents #12, HCT116 Pum1^−/−-2 represents #23, HCT116 Pum2^−/−-1 represents #10-4, HCT116 Pum2^−/−-2 represents #8-4, RKO Pum1^−/−-1 represents #3-4, RKO Pum1^−/−-2 represents #3-18, RKO Pum2^−/−-1 represents #2-19 and RKO Pum2^−/−-2 represents #2-27 according to numbers in Supplementary Fig. 5d, f. Tubulin was used as a loading control. d, e Representative images of colony formation assay in WT, Pum1^−/− and Pum2^−/− HCT116 (d, n = 2) and RKO (e, n = 3) cells. Bar graphs show the colony numbers per dish. Error bars represent SD. f, g Growth curves of WT, Pum1^−/− or Pum2^−/− HCT116 (f, n = 4) and RKO (g, n = 3) cells. Error bars represent SD. h, i Cell doubling time of WT, Pum1^−/− or Pum2^−/− HCT116 (h, n = 2) and RKO (I, n = 3) cells. Error bars represent SD. j, k Cell cycle analysis of WT, Pum1^−/− or Pum2^−/− HCT116 (j, n = 2) and RKO (k, n = 3) cells. Error bars represent SD. l–n Tumor growth (l), tumor volume (m), tumor weight (n) in mice injected with WT, Pum1^−/− or Pum2^−/− HCT116 cells (n = 8). Error bars represent SD. o–q Tumor growth (o), tumor volume (p), tumor weight (q) in mice injected with WT, Pum1^−/− or Pum2^−/− RKO cells (n = 8). Error bars represent SD. Source data are provided as a Source Data file.