Fig. 7: PP1 dephosphorylates HDAC4.
From: Crosstalk of HDAC4, PP1, and GSDMD in controlling pyroptosis
A Interaction of HDAC4 and PP1. IB of total cell lysates (input) and proteins immunoprecipitated with anti-Flag resin from HEK293T cells transfected with Flag-HDAC4 and HA-PP1. B, C Structural model of the complex of GSDMD, HDAC4 and PP1 (PDB: 4MOV). The docking models were analyzed using HDOCK Server. D Analysis of phosphorylation of HDAC4 in HEK293T cells transfected with Flag-GSDMD and HA-PP1. The cells were lysed, and their proteins were separated by Phos-tag PAGE and subjected to Western blot analysis. E THP-1 cells were treated with or without Okadaic acid (OA) and then lysed. Proteins were separated by Phos-tag PAGE and subjected to Western blot analysis. HEK293T cells were first transfected with HDAC4 and PP1. After 24 h, cells were then transfected with GSDMD-p30 (F) or GSDMD-FL and Caspase-1 (G) or GSDMD-FL and Caspase-4 (H). The supernatants were collected for LDH assay. I Immunofluorescence microscopy and nuclear staining (with the DNA-binding dye DAPI) of HEK293T cells transfected with expression plasmids for GFP-HDAC4, Flag-GSDMD and HA-PP1γ. Scale bars, 20 μm. ** stands for P < 0.01 and ns stands for no significant difference (unpaired t test). Data shown are mean ± SD from one representative experiments performed in triplicate.
