Fig. 4

IDH1/MDH1 deacetylation enhanced NETs formation in mice with LPS/d-gal-induced liver injury. A, B In dHL-60 cells, with plasmid mutations at the K118 site of MDH1 and the K93 site of IDH1, cells were stimulated with PMA in vitro, and the expression of CitH3 was measured by western blotting and analyzed by ImageLab. C, D In dHL-60 cells, the expression of PAD4 was measured by western blotting and analyzed by ImageLab (N: dHL-60; M: dHL-60 + PMA; IDH1-WT: IDH1-wild-type + PMA; IDH1-K93R: IDH1-K93R + PMA; IDH1-EV: IDH1-Flag + PMA; MDH1-WT: MDH1-wild-type + PMA; MDH1-K118R:MDH1-K118R + PMA; MDH1-EV: MDH1-HA + PMA.). E Liver samples were detected for NETs by immunofluorescence staining with MPO (green), CitH3 (red), and DAPI (blue); the scale bar is 50 μm. F The mean fluorescence intensity of Cit-H3 and MPO was quantified using ImageJ analysis software. Five microscopic fields per liver section were counted. N: liver samples injected with normal saline; M: ALF model, liver samples injected with LPS/d-gal; ACY1215: LPS/d-gal + ACY1215; IDHI-K93R: IDHI-K93R: adenovirus-infected mice + LPS/d-gal; MDHI-K118R: MDHI-K118R adenovirus-infected mice + LPS/d-gal group; EV: empty adenovirus-infected mice + LPS/d-gal, MPO Myeloperoxidase, CitH3 Citrullinated histone H3, DAPI 4′,6-diamidino-2-phenylindole, PAD4 peptidylarginine deiminase 4, ns no significance. The histograms show the means ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001