Figure 5. Smad4, Hmga2 and Notch1 are targets of miR-146b during myoblast differentiation.

(A) C2C12 cells were induced to differentiate, and total RNA was isolated from the differentiating cells on various days as indicated (Diff. day) and subjected to analysis by qRT-PCR to determine the relative levels of mRNA for each gene, with that at day 0 as 1. (B) Regeneration of mouse TA muscles was induced by BaCl₂ injury. On day 3 and day 5 AI, the TA muscles were isolated for RNA extraction, followed by qRT-PCR assays to measure relative mRNA levels. Saline injection into contralateral TA muscles served as control (“−”) and was designated as 1. (C) C2C12 cells were transfected with 50 nM LNA-anti-miR-146b or LNA control for 1 day and then induced to differentiate for 3 days. Total RNA was isolated on day 0 and day 3 of differentiation and analyzed by qRT-PCR. (D) Cells as described in C were lysed and subjected to Western blotting analysis. Data shown are mean ± SD from three to four independent experiments (A & C) or at least three mice per time point (B), or representative results of three independent experiments (D). One-sample t test was performed to analyze data in A & B, and paired t test was performed for data in C. *P<0.05; **P<0.01.