Fig. 5

HDAC3 deficiency in macrophages results in impaired RIP1-dependent NF-κB signaling activity. A, B Western blot analysis of phosphorylated and total RIP1 in Vector and Hdac3^−/− RAW264.7 stimulated with TNFα (20 ng/ml) for indicated time. C, D Western blot analysis of phosphorylated and total P65 in control and HDAC3 deficient RAW264.7 stimulated with TNFα (20 ng/ml) for indicated time. E, F Confocal micrographs of RAW264.7 stimulated or unstimulated with TNFα (20 ng/ml) for 15 min stained with DAPI (blue, DNA) and antibodies against P65 (AF488). The nuclear P65 positive cells were calculated. Scale bar: 100/10 μm. Ctl, Control. G PCR analysis of Il1β, Mcp1, Mip2 and Cox2 in Vector and Hdac3^−/− RAW264.7 stimulated with TNFα (20 ng/ml) for 1 h. Data are representative of three independent experiments and showed as mean ± SEM; *P < 0.05 and **P < 0.01