Figure 8

Involvement of HDAC1 in the inhibition of CD200R1 expression by C/EBPβ. (A) Western blot showing HDAC1 (upper image) and C/EBPβ (lower image) expression in nuclear protein extracts from primary mixed glial cultures before (Input) and after immunoprecipitation (IP) with anti-C/EBPβ antibody. HDAC1-C/EBPβ co-immunoprecipitation was observed in LPS-treated cultures. Samples immunoprecipitated with isotype or unlabeled beads were used as controls to demonstrate the specificity of the signal obtained. (B) Analysis of HDAC1 binding to C/EBPβ box 6 of CD200R1 gene promoter by qChIP, showing interaction between HDAC1 and C/EBPβ in LPS-treated cultures. Bars are means + SEM of three independent experiments. ***P <0.001 versus IgG and C. One-way ANOVA and Bonferroni post-test. Effect of the HDAC inhibitors SAHA (C) and MS-275 (D) on CD200R1 mRNA expression in control and LPS-treated primary mixed glial cultures. Both attenuated LPS-induced inhibition of CD200R1 expression. Bars are means + SEM of three independent experiments. *P <0.05, **P <0.01 and ***P <0.001 versus C; ##P <0.01 versus LPS. One-way ANOVA and Bonferroni post-test. ANOVA, analysis of variance; C, control; C/EBPβ, CCAAT/enhancer binding protein β; HDAC1, histone deacetylase 1; IgG, immunoglobulin G; LPS, lipopolysaccharide; qChIP, quantitative chromatin immunoprecipitation; SAHA, suberoylanilide hydroxamic acid; SEM, standard error of the mean.