nNOS depletion abolishes the neuroprotective effects of SU5416 against MPP+-induced neuronal death in PC12 cells.

Abstract

<p>(A) PC12 cells were transfected with pG418-GFP plasmid (vector), pG418-GFP plasmid encoding nNOS ShRNA (ShnNOS) and pG418-GFP plasmid encoding negative control ShRNA (ShNC). The levels of nNOS and β-actin in the cell lysates were analyzed by Western blotting assay by using specific antibodies. (B, C) nNOS depletion abolished the neuroprotective effects of SU5416 against MPP⁺-induced neuronal death in PC12 cells. PC12 cells transfected with vector, ShnNOS, or ShNC were treated with 20 µM SU5416 for 2 hours and then exposed to 1 mM MPP⁺. Cell viability (B) and cytotoxicity (C) were measured at 24 hours after MPP⁺ challenge by MTT and LDH assays, respectively. Data were the mean ± SEM of three separate experiments; **<i>p</i><0.01 <i>versus</i> control; ^##<i>p</i><0.01 <i>versus</i> MPP⁺ group; ^&<i>p</i><0.05 and ^&&<i>p</i><0.01 <i>versus</i> MPP⁺ vector group (Turkey’s test).</p