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2.7. Antibodies and reagents
The primary SGI1776 used in this study are as follows: anti-caspase-11 rat monoclonal antibody (1:200, Abcam), anti-PARP-1 mouse polyclonal antibody (1:500, BD Bioscience), anti-α-tubulin mouse monoclonal antibody (1:4000, Sigma). IFN-γ was purchased from R&D. 3,4-Dihydro-5-[4-(1-piperidinyl)butoxyl]-1(2H)-isoquinoline (DPQ) and 1,5-isoquinolinediol were purchased from Calbiochem. All other reagents were purchased from Sigma unless stated otherwise.
2.8. Statistics
For the statistical analysis, all the experiments were repeated at least three times. The results were expressed as means ± SD of at least three independent experiments, unless stated otherwise. Paired data were evaluated by ANOVA.
3. Results
3.1. Knockdown of PARP-1 suppressed the induction of caspase-11
Fig. 1.
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To determine whether PARP-1 is required for the transcriptional activation of caspase-11 gene, we examined the mRNA level of caspase-11 in PARP-1 knocked-down MEFs by RT-PCR at 2 h after LPS (1 μg/ml) stimulation. As shown in Fig. 1B, LPS-induced transcription of caspase-11 mRNA was also reduced by PARP-1 knockdown, suggesting that PARP-1 can regulate the induction of caspase-11 at a transcriptional level.