Fig. 3

eIF3a and eIF3b, the binding partners of METTL16, promote CSC self-renewal and HCC development. A, B Representative images (A) and the statistical results (B) showing the effects of KO of eIF3a or eIF3b on liver CSC maintenance as determined by spheroid formation assay in HepG2 cells (n = 4; mean ± SD). C, D Representative images (C) and the statistical results (D) showing the effects of KO of eIF3a or eIF3b on liver CSC maintenance as determined by spheroid formation assay in Hep3B cells (n = 3; mean ± SD). E, F Representative images (E) and statistical results (F) showing the effects of KO of eIF3a or eIF3b on liver CSC self-renewal ability as determined by in vitro LDA in HepG2 cells. G, H Representative images (G) and statistical results (H) showing the effects of KO of eIF3a or eIF3b on liver CSC self-renewal ability as determined by in vitro LDA in Hep3B cells. I, J Effect of KO of eIF3a or eIF3b on the population of liver CSCs (EpCAM⁺/CD133⁺) in HepG2 (I) and Hep3B (J) cells upon as determined by flow cytometry [n = 3 (I), n = 4 (J); mean ± SD]. K CERES scores of eIF3 subunits from genome-scale CRISPR–Cas9 essentiality screens across 23 liver cancer cell lines. The raw data were downloaded from DepMap (https://depmap.org/portal/). The lower CERES score indicates a higher cancer dependency of the specific gene. Each stick represents one HCC cell line. L Comparison of the mRNA levels of eIF3a and eIF3b between human HCC tissues and normal controls. Adjacent = 193, HCC = 240. The three lines inside the violin plots are the first quartile, median and third quartile. M, N Effect of eIF3a (M) and eIF3b (N) KO on proliferation of HepG2 cells (n = 5; mean ± SD). O eIF3a or eIF3b KO efficacy in the xenograft liver tumors implanted with HepG2 cells as determined by immunohistochemistry. P Average growth curves of xenograft liver tumors upon KO of eIF3a or eIF3b (n = 10). Q Weights of the liver tumors on day 28 post injection (n = 10; mean ± SD). R Kaplan–Meier disease-free survival (DFS) of the xenograft models implanted with HepG2 cells with or without KO of eIF3a or eIF3b (n = 10). Statistical analyses: unpaired t-test (B, D, I, J, L, Q); Log-rank test (R); Two-way ANOVA (M, N, P). **P < 0.01, ***P < 0.001