Figure 2

Knockdown of EZH1 and/or EZH2 in Huh7 and HepG2 cells. (A) Stable EZH1 knockdown cells were subjected to Western blot analyses. (B) Stable EZH1 and/or EZH2-knockdown cells were subjected to Western blot analyses. (C) Limiting dilution assay of Huh7 and HepG2 cells transduced with sh-EZH1, sh-EZH2 and sh-EZH1/2. The sphere formation frequency was calculated by extreme limiting dilution analysis (*p < 0.05, **p < 0.01). (D) Representative bright-field images of spheres formed on day 14 of low attachment cell culture. Fluorescence images are shown in insets. Scale bar = 100 µm. (E) Number of large spheres generated from 1000 cells transduced with the indicated viruses on culture day 14 (Student’s t test, *p < 0.05). (F) RNA sequence was performed using stable EZH1 or EZH2-knockdown Huh7 cells. Venn diagram depicting the number of genes showing de-repression after EZH1- or EZH2 knockdown.