Fig. 8
CtBP effects on p300 and NF-κB acetylation. a Chromatin immunoprecipitation (ChIP) with antibody to HDAC1, acetyl H3, and p65 was performed in RAW264.7 cells to evaluate binding to the IL-6 promoter regions. LPS increased all three signals, but only the effect of p65 binding was reversed by 2DG (conditions as in Fig. 1c); n = 3, *p < 0.05. b Western blots show LPS-induced p65 acetylation is suppressed by 2DG (conditions as in Fig. 1c); n = 3, *p < 0.05. Full length immunoblots are shown in Supplementary Fig. 3. c ChIP was performed to evaluate p300 binding to NF-κB p65 binding sites on promoter regions of pro-inflammatory cytokines. p300 binding was increased by LPS, and this effect was attenuated by 2DG (conditions as in Fig. 1c); n = 3, *p < 0.05. d HEK293 cells were transfected with FLAG-tagged WT CtBP2 or G189A CtBP2. Immunoprecipitation using antibody to FLAG recovered p300 protein, while antibody to IgG, used as a negative control, did not. Immunoprecipitates from transfected cells treated with 2DG or CoCl2 for 30 min showed reduced p300 binding to WT CtBP2, but not G189A CtBP2, in cells treated with CoCl2. e Quantified results of the immunoprecipitation studies. Results were normalized to p300 in the lysate input. n = 3; *p < 0.05. Error bars show s.e.m
