Fig. 2: Acetylation of GSDMD affects pyroptosis.
From: Crosstalk of HDAC4, PP1, and GSDMD in controlling pyroptosis
A Acetylation of exogenous Flag-GSDMD in HEK293T cells treated with deacetylase inhibitors TSA or NAM. Flag-GSDMD was immunoprecipitated with anti-acetyl-lys antibody (Ace-lys), and the precipitates were analyzed using an anti-flag antibody. B Acetylation of endogenous GSDMD in THP-1 cells. GSDMD acetylation was analyzed by immunoprecipitation with an anti-acetyl-lys antibody followed by western blotting. C Acetylation of exogenous Flag-GSDMD from pig or mouse in HEK293T cells. GSDMD acetylation was analyzed by immunoprecipitation with an anti-acetyl-lys antibody followed by western blotting. D, E Acetylation of endogenous GSDMD in IPEC-J2 (D), RAW264.7 (E) cells. GSDMD acetylation was analyzed by immunoprecipitation with an anti-acetyl-lys antibody followed by western blotting. F Acetylation of endogenous mouse-GSDMD from mouse lung. G–I HEK293T cells transfected with GSDMD-p30 or GSDMD-FL (full length) and Caspase-1 or GSDMD-FL and Caspase-4 were treated with TSA (10 μM). LDH release was analyzed using an LDH release assay. J–L THP-1 cells were incubated with 500 ng/mL LPS and TSA for 4 h and then another 1 h for 10 μM Nigericin. The supernatants were collected and analyzed by LDH release assay (J) and ELISA for IL-1β (L). Cell lysates were analyzed by immunoblotting (K). **** stands for P < 0.0001, *** stands for P < 0.001, ** stands for P < 0.01, * stands for P < 0.05 and ns stands for no significant difference (unpaired t test). Data shown are mean ± SD from one representative experiments performed in triplicate.
