Figure 3. Regulatory effects of miR-25 and miR-92a on insulin synthesis overrides that of miR-9 in the downstream insulin secretion. Changes in insulin protein in INS-1 cells with the corresponding transfection as shown above followed by high glucose treatment (20 mM). In the left panel, cells were first transfected with anti-miR-9 followed by independent transfection of anti or pre miR-25 or miR-92a . Cells transfected with anti-miR-9 and corresponding scrambled miRNAs (anti or pre) followed by the same high glucose (20 mM) treatment were used as controls). In the right panel, cells were first transfected with pre miR-9 followed by independent transfection of anti or pre miR-25 or miR-92a. Cells transfected with pre miR-9 and corresponding scrambled miRNAs (anti or pre) followed by the same high glucose (20 mM) treatment were used as controls. All transfections of miRNAs were done at 30 nM concentration. Positive sign (+) indicates presence of corresponding anti- or pre-miRNAs while negative sign (–) indicates absence. Data presented as mean ± SEM (n = 3) against corresponding controls. Intracellular insulin content (in white bar) and secreted insulin (in black bar) are normalized to total protein content. Statistically significant differences are tested using t-test at *p < 0.05 significance.