Anti n wasp

Liver tissues were fixed in 4% neutral buffered formalin and embedded in paraffin. The paraffin sections were deparaffinized by baking and then dehydrated using xylene and ethanol. After blocking with 1% bovine serum albumin (BSA) for 1 h, the sections were incubated with anti-CD11b (1:100, BD Biosciences, USA), anti-Ly6G (1:100; BD Biosciences), anti-H3cit (1:400; CST, USA), anti-MPO (1:100; Abcam, UK), and anti-N-WASP (1:100; Abcam) antibodies overnight at 4°C. Cells were fixed in 4% paraformaldehyde for 25 min at room temperature and then permeabilized with 0.2% Triton X-100 in PBS for 10 min. The cells were blocked in PBS with 2% BSA for 30 min at room temperature and then incubated with anti-H3Cit (1:400; CST), anti-N-WASP (1:100; Abcam), or anti-MPO (1:100; Abcam) antibodies overnight at 4°C. The sections and cells were then incubated with the indicated Alexa Fluor-conjugated secondary antibodies. DAPI was used to detect nuclei. Between all steps, samples were washed three times in PBS for 5 min each. The sections were visualized using an LSM 800 laser scanning confocal microscope (Zeiss, Germany).