Fig. 3
From: The protective role of DOT1L in UV-induced melanomagenesis
DOT1L mutations cooperate with BRAFV600E to promote melanomagenesis. a Engineered human melanocytes were infected with the shDOT1L lentiviral construct to deplete endogenous DOT1L, with control or BRAFV600E expression. b The cells generated in a were subjected to clonogenic survival assays 14 days after UVR. Crystal violet was used to stain the colonies and the relative colony formation was calculated as mean ± s.d. c The cells generated in a were seeded (10,000 cells per well) for soft agar assay and cultured for 30 days. The relative colony numbers were calculated as mean ± s.d. d DOT1L-depleted hTERT/p53DD/CDK4(R24C)/BRAFV600E melanocytes expressing empty vector (EV), DOT1L wild-type (WT), M55L, P271L, and P505L mutants were subjected to western blot. e The cells generated in d were subjected to clonogenic survival assays 14 days after UVR. Crystal violet was used to stain the colonies and the relative colony formation was calculated as mean ± s.d. f The cells generated in d were seeded (10,000 cells per well) for soft agar assay and cultured for 30 days. The relative colony numbers were calculated as mean ± s.d. g–i Growth curves for the xenograft experiments. The indicated 3 × 106 tumor cells were inoculated subcutaneously into each flank of the nude mice. The visible tumors were measured at the indicated days, error bars represent ± s.e.m, n = 7 (g). Dissected tumors (h) and tumor weight (h) were displayed. **p < 0.01, ***p < 0.001, unpaired Student’s t-test
