Fig. 4

MiR-101 forced expression as well as EZH2 pharmacological inhibition reduces eRMS cell migration. RD (a) and JR1 (c) cells were infected with pS-pre-miR-101 or control pS- retrovirus. Twenty-four hours post-infection cells were seeded on inserts and lived to reach the confluence for 48 h, when the inserts were removed. Cells were imaged at 0 and 24 h or 36 h after the insert removal. RD (b) and JR1 (d) were treated with DZNep (5 μM) or vehicle (i.e., water, referred as untreated condition: UN) for 72 h and then inserts were removed and cells were imaged as in (a) and (b). Representative phase contrast microscopy images of the migration assays at 0 and 24 h or 36 h after gap creation were shown. Dashed lines indicate the boundary of the edges of the wound at 0 and 24 h or 36 h. The histograms depict the measurements of the total area between the wound edges of the scratch from at least five random fields per scratch from two separate experiments, expressed as fold change over control pS- (a and c, 1 arbitrary unit) or untreated (UN) (b and d, 1 arbitrary unit) samples. Columns, means; bars, SD. *P < 0.05 (Student’s t-test)