Figure 6.

circSNX29 Binding miR-744 to Promote Differentiation of Bovine Primary Myocytes

(A) circSNX29 expression in different stages of differentiation. (B–K) Myoblasts were treated with pcDNA2.1-circSNX29 (or) pcDNA3.1-miR-744 and pcDNA2.1-circSNX29. (B and C) The mRNA and protein expression of cell differentiation markers (MyoG and MyoD) were measured by real-time qPCR (B) and western blotting (C). (D) Cell differentiation was also measured by immunofluorescence assay; the scale bars represent 200 μm. Cell phase was analyzed (E) and counted (F) by flow cytometry. (G) Cell proliferation index was detected by CCK-8 assay. (H) Cell proliferation was detected via MTT assay. (I) Cell proliferation was detected with EdU; the scale bars represent 100 μm. (J and K) The expression of proliferation marker gene PCNA and cyclin D1 was measured by real-time qPCR (J) and western blotting (K). Values are mean ± SEM for three biological replicates; *p < 0.05; **p < 0.01.