Figure 3.

miR-590 Directly Targets Acvr2a to Regulate Activin Signaling in mESCs

(A) Summary of miR-590-3p and miR-590-5p target sites in the 3′UTRs of Acvr2a. In the double strands of the sequence, the upper strand is miR-590-3p or miR-590-5p, and the lower strand is the binding sites of 3′UTR mRNA. The right panel shows the level of Acvr2a expression detected by qRT-PCR. Data shown are means ± SD of three independent experiments (n = 3). ^∗p < 0.05, ^∗∗p < 0.01, and ^∗∗∗p < 0.001.

(B) miR-590-3p and miR-590-5p specifically repress their targets in the luciferase assay. WT-UTR indicates WT 3′ UTR. Mut-UTR indicates the 3′UTR containing the mutant binding site of miR-590. Data shown are means ± SD of three independent experiments (n = 3). ^∗∗∗p < 0.001.

(C) Detection of Acvr2a expression on mRNA and protein levels in mESCs. Data shown are means ± SD of three independent experiments (n = 3). ^∗p < 0.05.

(D and E) The level of p-SMAD2 was detected by western blot analysis. Glyceraldehyde-3-phosphate dehydrogenase is as an internal control.