Figure 4
miR-486-5p is a downstream effector of PAX3-FOXO1 exosome-mediated effects. (a,b) qRT-PCR analysis of miR-486-5p expression in (a) P3F-C2C12 cells and (b) their derived exosomes, relative to Ctrl-C2C12 cells and their derived exosomes, respectively, using 2 distinct internal controls, let-7a and snoRNA202 (c) Relative expression of miR-486-5p known targets that were significantly downregulated in MEFs treated for 48 h with P3F-C2C12 exosomes normalized to those treated with Ctrl-C2C12 exosomes as detected by qRT-PCR.GAPDH was used as internal control. (d,e) Migration (d) and invasion (e) of MEFs treated with 10X exosomes extracted from P3F-C2C12 cells transduced either with negative control (Scrm), or with knockdown of miR-486-5p by MiRZip-486 (Mzp-486-5p), as indicated. Histograms represent ratio of the denoted conditions versus control. (f) Ratio of number of colonies formed by P3F-C2C12 cells after treatment with the specified exosomes at 10X, to that of cells treated with control exosome-free media. Representative microscope images are of colonies at 3 weeks. Values are mean ratio of three independent experiments. Bars represent standard deviation. Asterisks denote a statistically significant difference (p-value < 0.05).
