Figure 5

Effect of miR-7a on UPR-mediated cell death. (A) Upper panel, Shows a schematic representation of Lentiviral vector used to generate miR-7a expressing clones. Lower panel, H9c2-control and H9c2-miR-7a cells were treated with (1 μg/ml) of doxycycline for 24 h and expression levels of miR-7a was quantified by qRT-PCR, normalizing against snoRNA. Error bars represent mean ± S.D. from two independent experiments performed in triplicate. (*P < 0.05, two-tailed unpaired t-test as compared with uninduced cells). (B) Expression of GFP was monitored in H9c2-control and H9c2-miR-7a cells after treatment with (1 μg/ml) of doxycycline for 24 hours. (C) The H9c2-control and H9c2-miR-7a cells were treated (2 μg/ml) Tm for the indicated time. Histograms of annexinV-PE binding as obtained by FACS analysis of a representative experiment are shown. Numbers depict the percentage of annexin-positive cells. (D) The H9c2-control and H9c2-miR-7a cells were treated with (1 μM) Tg and (2 μg/ml) Tm for the indicated time and (1 μM) staurosporine for 24 hours. Western blotting of total protein was performed using antibodies against cleaved caspase-3 and actin.