Fig. 6

CircUBE2Q2 acts as a sponge for miR-133a. A RT-qPCR analysis of miR-133a levels in MuSCs transfected with circUBE2Q2. B miR-133a binding sites in the 3′-untranslated region (UTR) of circUBE2Q2 obtained by bioinformatics. C Luciferase assay of MuSCs transfected with the psiCHECK2 vector (control) and pCK-circUBE2Q2 reporter with a miR-133a mimic (n = 3). D Fluorescent in situ hybridization assay shows circUBE2Q2 (red), miR-133a [28], and nuclei (DAPI, blue) in MuSCs. Magnification 73x. E MuSCs were transfected with miR-133a and circUBE2Q2, and cell proliferation was measured with 5-ethynyl-20-deoxyuridine (EdU). F EdU-positive cell index statistics are shown. G Cell proliferation was assessed using the cell counting kit-8 (CCK-8) assay. H The expression of the proliferation marker gene, PCNA, was measured using RT-qPCR. I MuSCs were transfected with the miR-133a and circUBE2Q2, and immunofluorescence detected cell differentiation. J and K The expression of the myocyte differentiation marker genes MyOD1 (J) and MyHC (K) were detected by RT-qPCR. Values are means ± SEM. Each letter denotes a significant difference (P < 0.05). Scale bars = 100/400 μm