Fig. 1

EZH2 is overexpressed in UM. a-b The mRNA (a) and protein (b) levels of EZH2 in normal choroid of healthy donors (3 for qRT-PCR, 2 for Western blotting analysis), UM tissues (n = 3) and UM cells (92.1, Mel270, Omm2.3 and Omm1). Each dot represents one sample or one cell line. Data are mean ± SEM. **, P < 0.01, one-way ANOVA, post hoc intergroup comparisons. c-e IHC staining of EZH2 in paraffin embedded UM tissues. Choroid was adjacent normal tissue. H&E staining and IHC analysis of HMB45 were the same samples as those for EZH2 staining. ♦, sclera; ▲, retina. Scale bar, 100 μm, Olympus IX71 (c). The percentage of UM patients was divided by scores of EZH2 expression (d). Data are mean ± SEM. ***, P < 0.0001, Student’s t test (e). f-h E2F1 conferred EZH2 overexpression in UM. E2F1 expression was detected by qRT-PCR in normal choroid of healthy donors (n = 3), UM tissues (n = 3) and UM cells (n = 4). Each dot represents one sample or one cell line. Data are mean ± SEM. **, P < 0.01, one-way ANOVA, post hoc intergroup comparisons (f). Ectopic expression of E2F1 in Mel270 led to aberrant expression of EZH2 as detected by qRT-PCR (g) and Western blotting analysis (h). Data are mean ± SEM. **, P < 0.01, Student’s t test. i-l EZH2 promoted proliferation of UM cells. UM cells transfected with vector, EZH2-encoding constructs (i), scramble or lentiviral shRNAs against EZH2 with or without EZH2 restored (j-l) were subjected to either cellular growth determination (i and k), colony formation evaluation (j) or Western blotting analysis (l). Data are mean ± SEM. **, P < 0.01; ***, P < 0.0001, one-way ANOVA, post hoc intergroup comparisons