Fig. 7
Direct inhibition of CtBP dimerization blocks LPS-induced pro-inflammatory gene expression. a Schematic of CtBP1 protein showing functional domains and alignment with the CtBP peptide used to block CtBP dimerization. PLDLS indicates substrate binding domain. The CtBP blocking peptide includes an N-terminal TAT sequence for cellular internalization. b Immunoprecipitation assay showing ability of CtBP peptide (50 µm) to block dimerization of tagged CtBP proteins incubated with 10 mM lactate + 10 µm NADH. Full length immunoblots are shown in Supplementary Fig. 3. Immunoprecipitation was performed with anti-Flag antibody, and CtBP1-Flag/CtBP1-HA heterodimers were detected by western blots using anti-HA antibody. Lysate control lane = 5% of input used in immunoprecipitated samples. n = 4. **p < 0.01. c CtBP peptide (5 µm) blocks LPS-induced mRNA expression of pro-inflammatory genes (iNOS, IL-1b and IL-6) in cultured primary microglia. Ctrl PEP = control peptide. n ≥ 3; **p < 0.01. d 2DG blocks LPS-induced iNOS gene expression in microglia isolated from mice after LPS (8 µg) injection into striatum. n = 3–5 per group. **p < 0.01. e, f CtBP peptide blocks LPS-induced iNOS expression but not Socs3 expression in brain microglia. n = 4 per group. **p < 0.01. Error bars show s.e.m
