Upregulation of miR-7a expression in primary cardiomyocytes during ischemia. (A) H9c2 cardiomyocytes were either untreated (Control) or treated with (1.0 μM) Tg and (1.0 μg/ml) Tm for indicated time points. The expression level of indicated miRNAs was quantified by qRT-PCR, normalizing against snoRNA. Error bars represent mean ± S.D. from three independent experiments performed in triplicate. (B) H9c2 cells were treated with 2-deoxyglucose (1 mM) along with serum and glucose deprivation for 24 hours. The change in expression levels of ER stress markers normalized against GAPDH expression and miR-7a normalized against snoRNA expression was measured by qRT-PCR. The expression levels relative to the control are shown. SFM, serum free medium; GFM, glucose free medium and 2DG, 2-deoxyglucose. (C-D) Cardiomyocytes were exposed to simulated in vitro ischemia (Ischemia) consisting of glucose-free anoxia at pH 6.4 for 3 hours and cardiomyocytes not exposed to ischemia were used as controls (Control). The change in expression levels of ER stress markers normalizing against GAPDH expression (C) and miR-7a PCR normalizing against snoRNA expression (D) was measured by qRT-PCR. The expression levels relative to the control are shown. (*P < 0.05, two-tailed unpaired t-test compared with untreated cells).