Figure 2

c-Myb suppression inhibits colony formation of eRMS cell line RD but not aRMS cell line RH30. (a) RD or (b) RH30 cells transduced with a Dox-inducible c-Myb shRNA (RD^shMYBor RH30^shMYB) vector or empty vector (RD^Emptyor RH30^Empty) were cultured with or without Dox (5 μg/ml for RD cells and 2.5 μg/ml for RH30 cells) in 0.35% agar for 21 days. Formed colonies were fixed, stained, the size of colonies was determined, and density plot was created from the log cell area. It is shown that cells form colonies of various sizes: small colonies with a countable number of cells (peaks) are divided from larger colonies formed by an uncountable number of cells by a black line. Higher density shows more cells within the same area.