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Fig. 6 | Journal of Neuroinflammation

Fig. 6

From: NF-κB transcriptional activation by TNFα requires phospholipase C, extracellular signal-regulated kinase 2 and poly(ADP-ribose) polymerase-1

Fig. 6

TNFα-induced PARP activation is regulated by ERK2. a TNFα (15 ng/ml) induces ERK1/2 phosphorylation in astrocyte cultures. Representative of n = 3. b PAR immunoblots shows that TNFα-induced PAR formation is reduced after siRNA downregulation of ERK2, but not ERK1. PAR is seen as a discrete band at 116 kDa attributable to auto-poly(ADP)-ribosylation of PARP-1 and as a smear over other molecular weights resulting from PAR formation on other proteins. Actin band shows protein loading. ERK1 and ERK2 expression are shown below the PAR western blot. The siRNA treatments reduced their expression by 72 ± 8 % and 72 ± 5 %, respectively, with protein loading shown by the actin band. c Quantification of PAR western blots, conditions as in b; * p < 0.05, n = 3. d Immunostaining for PAR also shows that TNFα-induced PAR formation is suppressed by siRNA downregulation of ERK2 expression. Images are representative of four independent experiments. All PAR formation studies are conducted 60 min after TNFα stimuli

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