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. 2013 Mar 20;8(3):e58697. doi: 10.1371/journal.pone.0058697

Figure 8. Knockdown of GAPDH attenuates Mst1 activation and cell apoptosis in response to chelerythrine.

Figure 8

A, Total RNA extracted from control siRNA (siCTL) and GAPDH siRNA transfected cells was analyzed for the expression of GAPDH in NRCMs by qRT-PCR. B, NRVMs were transfected with either control siRNA or GAPDH siRNA. 72 hours after transfection, cell lysates were then subjected to western blot analysis to detect the expression of GAPDH. C, NRVMs were transfected with either control siRNA or GAPDH siRNA. 72 hours after transfection, cells were treated with chelerythrine (5 µM) for 2 hours. Mst1 was then immunoprecipitated and its activity was determined by an in vitro kinase assay using histone H2B as a substrate. D, NRVMs were transfected with either control siRNA or GAPDH siRNA. 24 hours after siRNA transfection, cells were then transduced with either Ad-LacZ or Ad-Mst1 (MOI = 50). 48 hours after virus transduction, NRVMs were treated with chelerythrine (5 µM) for 2 hours and the cell apoptosis was determined by using the TUNEL staining kit (Roche). Values are means ± SEM obtained from 4 experiments.