Fig. 2: m6A factors are altered in RMS and sustain its proliferation and migration rate.
a Protein levels for several components of the MACOM complex, as well as for the reader YTHDC1, in orthotopic RMS patient-derived xenografts, compared to normal myoblasts and myotubes. Data derive from https://pecan.stjude.cloud/proteinpaint/study/RHB2018. A vertical dashed line indicates the level of protein expression of each gene in normal myoblasts. b Representative western blot to evaluate the levels of METTL3, METTL14, and YTHDC1 in wild-type myoblasts, RD and RH4 cell lines; GAPDH was used as loading control. n = 3 biologically independent replicates. c Relative number of cells upon control treatment (“si-scr”) or METTL3 depletion (“si-METTL3”) in RD or RH4 cell lines 48 hrs post transfection. Data are represented as mean percentage of cells ± SD. n = 4 biologically independent replicates. d Cell cycle analysis by FACS of RD or RH4 cells either upon control treatment (“si-scr”), METTL3 knock-down (“si- METTL3”), or METTL14 knock-down (“si- METTL14”). Data are represented as mean percentage of cells in each cell cycle phase ± SD. n = 4 biologically independent replicates. e Relative number of cells/field measured after transwell-migration assay with DAPI staining upon control treatment (“si-scr”) or METTL3 depletion (“si-METTL3”) in RD and RH4 cells. Data are represented as mean percentage of migrated cells ± SEM. n = 4 biologically independent replicates. Where statistical analysis was performed, the ratio of each sample versus its experimental control was tested by a two-tailed unpaired Student’s t test (c, e) with correction for multiple test comparison (FDR Benjamini–Hochberg) (d). * indicates a test-derived p value < 0.05, ** indicate a p value < 0.01, and *** a p value < 0.001. Source data are provided as a Source Data file.
