Fig. 6: mROS inhibition or GSDMD knockout suppressed NETs in bone marrow neutrophils after CTSC treatment.

a Immunofluorescence staining of cit-H3 (Red), Sytox Green (Green), Elastase (magenta) and Hoechst (Blue) in peripheral blood neutrophils from healthy volunteers. Cells were treated with DMSO, CTSC, CTSC + MitoTEMPO (MT) or CTSC + LDC7559. Scale bar, 100 μm. b Quantification of the percentage of released DNA per field of view from indicated groups in (a). c Immunofluorescence staining of cit-H3 (Red), Sytox Green (Green), Elastase (magenta) and Hoechst (Blue) in bone marrow neutrophils from WT and Gsdmd^−/− mice. Cells were treated with DMSO or CTSC. Scale bar, 30 μm. d Quantification of the percentage of released DNA per field of view from indicated groups in (c). e Representative histograms of Mito-Sox in peripheral blood neutrophils from healthy volunteers. Cells were treated with DMSO, CTSC or CTSC+LDC7559. f Quantification of Mito-Sox intensity from indicated groups in (e). g Representative histograms of Mito-Sox in bone marrow neutrophils from WT or Gsdmd^−/− mice. Cells were treated with DMSO, CTSC or CTSC + LDC7559. h Quantification of Mito-Sox intensity from indicated groups in (g). i Quantification of LDH released into the supernatant from bone marrow neutrophils after DMSO, CTSC or CTSC with MitoTEMPO pretreatment. Data are presented as means ± SEM. Significance was examined with one-way ANOVA (b, d, f, h), two-tailed unpaired Student’s t test (i). MFI mean fluorescence intensity. ^*P < 0.05, ^***P < 0.001.