Figure 8
The protective effect of osmotin on ethanol-induced apoptosis requires adiponectin receptors. (a) Downregulation of adiponectin receptor expression by transfection with siRNA. Shown is the RT-PCR analysis of AdipoR1 and AdipoR2 mRNA expression in primary fetal (GD 17.5) hippocampal neurons that were transfected with unrelated siRNA or AdipoR1 plus AdipoR2 (1 : 1) siRNA. GAPDH mRNA levels are shown for normalizing expression levels. (b–d) Primary cultures of fetal hippocampal neurons that were transfected with a mixture (1 : 1) of AdipoR1 and AdipoR2 (AdipoR1+R2) siRNAs and unrelated siRNA were treated with ethanol (EtOH; 100 mM), osmotin (Osm; 0.16 μM), or adiponectin (Adipo; 0.1 μM) in combinations for 24 h. Cleaved PARP-1, cleaved capsase-3, and p-AMPK levels were then analyzed in extracts of the cultured hippocampal neurons by western blotting. Shown are the immunoblots for detection of cleaved PARP-1 (b), cleaved caspase-3 (c), p-AMPK (d), and β-actin as loading control in all cases. The graph represents normalized density values for the immunoblots signals expressed as mean±S.D. (n=4). Sample pairs that are significantly different from one another at P<0.05 are indicated
