Figure 5.

let-7b-5p Decreased IRS1 Expression in Cytosol

(A) Lipid deposition in miR-92a-2-5p- and let-7b-5p-treated H9c2 cardiomyocytes was detected by BODIPY probe. n = 3; *p < 0.05 versus control. (B) Pathway analysis was used to explore the function of genes potentially targeted to miR-92a-2-5p and let-7b-5p by DIANA-miRPath database. (C) mRNA in association with Ago2 in H9c2 cells was determined by quantitative real-time PCR. n = 3; results from control were set to 1. (D and E) Effects of transfected miR-92a-2-5p and let-7b-5p on IRS1, AKT, and IGF1 at the protein levels were detected by western blot (D) and were repeated three times independently (E). *p < 0.05 versus control. (F) Lipid deposition in si-IRS1-treated H9c2 cardiomyocytes was detected by BODIPY probe. n = 3. (G) Effects of miR-92a-2-5p and let-7b-5p on Cytb and IRS1 protein expression were detected by BODIPY probe with the miRNA machinery selectively inactivated by knocking down GW182 in the cytoplasm; n = 3; results from si-NC-random were set to 1. (H) Effects of miR-92a-2-5p and let-7b-5p on lipid deposition were detected by BODIPY probe with the miRNA machinery selectively inactivated by knocking down GW182 in the cytoplasm. Cells were incubated with palmitate (0.25 mmol/L) for 24 h. n = 3; *p < 0.05 versus si-NC-random; ^#p < 0.05 versus si-GW182-random.