Fig. 3: Expression of miR-29c and ASB2 in myogenic cells.

a qPCR analysis of miR-29c in RISC complexes derived from two CTR and two DM1 cell lines allowed to differentiate for 5 days. Levels of miR-29c in both INPUT and IP were normalized on miR-181a levels, and shown as fold change of DM1 vs CTR, referred as 1 (n = 6; *P < 0.05). b Expression of miR29c and ASB2 in CTR and DM1 undifferentiated myogenic cells (T0) and myogenic cells induced to differentiate for 3 and 5 days (T1 and T2, respectively). Expression of miR-29c and ASB2 mRNA was analyzed by qPCR (top and middle charts), normalized for expression of miR-16 and RPL23, respectively, while ASB2 protein expression was measured by western blot (bottom chart and gel image of a representative experiment), normalized to vinculin (VCL) expression. DM1 values were first normalized to CTR, referred as 1, for each experiment and then the average value and standard error of DM1 fold change was calculated (n ≥ 4; *P < 0.05; **P < 0.01; ***P < 0.001). c Expression of miR-29c and ASB2 protein in CRISPR/Cas9 treated clones, either maintaining CTG expansions (CTG+, average values of 2 clones) or with full deletion of repeat expansions (CTG−, average values of 2 clones), induced to differentiate for 2 days. Values were normalized for expression of miR-16 and vinculin, respectively, and shown as fold change of CTG− clones relative to CTG+ clones referred as 1. (n ≥ 3; *P < 0.05; ***P < 0.001). A representative western blot analysis of ASB2 protein in CRISPR-Cas9-edited clones and in untreated DM1 and CTR cells is shown