ATF3 Protects against LPS-Induced Inflammation in Mice via Inhibiting HMGB1 Expression
Figure 5
Effects of ATF3 knockout on LPS-induced expression of IL-6, TNF-, and iNOS and translocation of NF-κB p65 in mice and in macrophages. (a) Quantitative RT-PCR (mRNA, top panels) and ELISAs (protein, lower panels) of Il-6 and TNF- from the lung tissues of WT and ATF3−/− mice treated with LPS (5 mg/kg, ip). Expression of mRNA was normalized to that of glyceraldehyde 3-phosphate dehydrogenase. Data represent mean ± SEM (). *, , #, and §, ; NS indicates not significant. (b) shows plasma nitrite concentrations. Data represent mean SEM (). #, . (c) shows a representative Western blots of iNOS. The intensity of iNOS was normalized to that of β-actin which was served as the loading controls. Data represent mean ± SEM (). #, . (d) Representative Western blots of nuclear NF-κB p65 protein in the lung tissues of WT and KO mice after LPS treatment. HDAC1 was served as the internal standard. Data represent mean ± SEM (). *, . (e) Nuclear NF-κB p65 levels in RAW264.7 macrophages measured by Western blot analysis. The intensity of nuclear p65 was normalized to that of HDAC1. Nuclear NF-κB DNA binding activity in RAW264.7 macrophages measured by ELISAs in RAW264.7 macrophages after incubation with LPS (200 ng/mL) for 24 hrs. Data represent mean ± SEM (). *, .
