Figure 5. Isoproteronol and miR-199a-5p abrogate insulin-induced upregulation of Hif-1α in cardiac myocytes.

a. Cardiac myocytes were treated with insulin using various doses and durations, as indicated. The Hif-1α signal from 4 different experiments were quantified, normalized to GAPDH, averaged, and plotted relative to its control levels vs. insulin dose and stimulation duration. Error bars represent SEM. b. Cardiac myocytes were treated with insulin for various time intervals or Ad. miR.199a-5p for 24 h, following pretreatment with vehicle or 10 µM ISO for 16 h, as indicated (n=3). c. Myocytes were treated with 10 µM ISO for various time intervals before stimulating them with 200 nM insulin for 24 h (n=4). d. Cells were treated with Ad.miR-199a-5p or a control virus for 20 h followed by 200 nM insulin for 6 h, as indicated by the + sign. At the conclusion of all the treatments listed above, protein was extracted and analyzed by Western blotting for the molecules indicated on the left of each panel (n=3). e. Hif-1α signal from experiments shown in c and d were quantified, normalized to GAPDH, averaged, and plotted relative to its control levels after adjusting it to 1. Error bars represent SEM and *p<0.001 vs. control.