Fig 2: The ANCA-associated vasculitis model of mouse induced by human ANCA from patients. (A) The mouse model of ANCA-associated vasculitis, constructed with human IgG from healthy donors, anti-PR3 or anti-MPO vasculitis patients after neutrophil mobilization by LPS. (B) The representative pictures of the lung, kidney and spleen from all groups. (C, D) The comparison of urea nitrogen (C) and creatinine (D) in mouse serum. (E) The representative pictures of urine smear across all groups. (F, G) The comparison of hematuria (F) and albuminuria (G). (H, I) H&E and PAS staining images of the kidney (H) and H&E staining images of the lung (I) from mice of all groups. Scale bar, 20 µm. (J) The summarized number of crescents among all groups. (K) The detection of biochemical indexes in mouse serum. (L) The indirect immunofluorescence microscopy assay for mouse ANCA on mouse neutrophils. Scale bar, 10 µm. **P < 0.01, ***P < 0.001, ****P < 0.0001.

Fig 3: Correlation of CPT2 expression and immune infiltration in human specimens. (A, B) Correlation of CPT2 expression with Kaplan–Meier curves and clinical stages in COAD patients. (C) Relative mRNA expression of CPT2 in 28 paired human specimens, N normal vs. T tumor. (D) Representative multiplexed IHC image of tissue area (2 mm × 1.5 mm) with an enlarged image for COAD human specimens, showing staining of CPT2 (brown) and MPO (red). (E) Spearman correlation analyses suggested that the expression of CPT2 negatively correlated with that of MPO (Spearman r = -0.42, p < 0.01). Scale bars are 20 and 200 µm in (D).

Fig 4: Representative images of immunofluorescence staining of low (upper panel) and high (lower panel) NET density in cervical cancer samples. Magnification 4×, scale bar = 100 µm; magnification 20×, scale bar = 100 µm. MPO, Red; H3Cit, Green; DAPI, Blue.

Fig 5: HBV-mediated S100A9 facilitates NETs generation. (A) Procedure for NETs analysis by co-culture of neutrophils with QSG-7701, HepG2 or HepG2.2.15 cells for 48 h. (B) The representative images of IF staining for DNA/NE/MPO in neutrophils co-cultured with QSG-7701, HepG2 or HepG2.2.15 cells for 48 h. White scale bars, 50 µm. (C) Western blot analysis for CitH3 in neutrophils co-cultured with QSG-7701, HepG2 or HepG2.2.15 cells for 48 h. The fold change of densitometric ratios was normalized to the ß-actin and then compared to the control and was shown on the right panel. (D) ELISA analysis for MPO-DNA in cultured supernatant of co-culture system. (E) Western blot analysis for endogenic expression of S100A9 in QSG-7701, HepG2 and HepG2.2.15 cells. The fold change of densitometric ratios was normalized to the ß-actin and then compared to the control and was shown on the right panel. (F) ELISA analysis for S100A9 levels in cultured supernatant of QSG-7701, HepG2 and HepG2.2.15 cells. (G) ELISA analysis for S100A9 levels in cultured supernatant of HepG2.2.15 cells transfected with siNC or siS100A9 for 48 h. (H) The representative images of IF staining for DNA/NE/MPO in neutrophils treated with CM (siNC or siS100A9-transfected HepG2.2.15) for 48 h. White scale bars, 50 µm. (I) Western blot analysis for CitH3 in neutrophils treated with CM (siNC or siS100A9-transfected HepG2.2.15) for 48h. The fold change of densitometric ratios was normalized to the ß-actin and then compared to the control and was shown on the right panel. (J) ELISA analysis for MPO-DNA in cultured supernatant of neutrophils treated with CM (siNC or siS100A9-transfected HepG2.2.15) for 48 h. (K) The representative images of IF staining for DNA/NE/MPO in neutrophils treated with and without GST, GST-S100A9 or PMA for 12 h. White scale bars, 50 µm. (L) Western blot analysis for CitH3 in neutrophils treated with and without GST, GST-S100A9 or PMA for 12 h. The fold change of densitometric ratios was normalized to the ß-actin and then compared to the control and was shown on the right panel. (M) ELISA analysis for MPO-DNA in cultured supernatant of neutrophils treated with and without GST, GST-S100A9 or PMA for 12 h. None, no treatment. *P < 0.05, **P < 0.01, ***P < 0.001. Abbreviations: siS100A9, the siRNA to interfere with S100A9; siNC, negative control siRNA; GST, glutathione-s-transferase.