Anti cd163

Clinical nasopharyngeal carcinoma samples and C57 BL/6 tumor xenografts were analyzed by the histochemical stain. Briefly, the paraffin-embedded sections of tumor tissues were deparaffinized and rehydrated. After antigen retrieval with 0.1M citrate buffer (pH 6.0), sections were blocked with 5% BSA for 30 minutes. The tumor samples were incubated with primary antibodies at 4°C overnight. Anti-EBV ZEBRA (BZLF1) (#sc-53904, Santa Cruz), anti-EBV EBNA1 (ab20870, Abcam), anti-CD34(#ZA-0550, ZSGB-bio), anti-CD163(#GB113152, Servicebio), anti-CD1a (#MA5-12526, Invitrogen) were used as the primary antibodies. PAS staining was performed using a PAS Staining Kit (#G1008, Servicebio). Images were acquired using Zeiss observer Z1, the numbers of classic angiogenesis were counted from three randomly chosen fields. Quantification of the positive area of CD1a versus CD163 was performed by the IHC Profiler function of IMAGE J.

Immunofluorescence staining was performed on both lung tissue and cellular slices through routine operation. For tissue staining, after dewaxing, rehydrating, antigen repairing, and blocking, paraffin-embedded tissue slices were incubated with diluted primary antibodies of anti-F4/80 (#GB11027, Servicebio), anti-Cytokeratin 7 (CK7, #GB11225, Servicebio), and/or anti-ADAR1 (#SC-73408, Santa Cruz), anti-cleaved-GSDMD (#AF4013, Affinity, Jiangsu, China), TUNEL (#GDP1043, Servicebio), anti-iNOS (#GB11119, Servicebio), or anti-CD163 (#GB11340-1, Servicebio) at 4 °C overnight, followed by incubation with their corresponding secondary antibodies at room temperature for 1 h in the dark. Between double staining procedures, 488-TSA (Servicebio) incubation and reantigen repair were conducted. DAPI (#G1012, Servicebio) was used for nuclear counterstaining. The slices were mounted with antifluorescence quenching sealing reagents (Boster, Wuhan, China) and captured under a fluorescence microscope (Life Technologies, Carlsbad, USA). For cellular staining, the primary antibodies used were anti-cleaved GSDMD (#AF4013, Affinity) and anti-caspase-1 (#GB11383, Servicebio). Mean fluorescence intensity (MFI) in a random field of view was measured using ImageJ software.

Liver tissues were cut into 5-µm thickness slides and incubated with anti-CD68 (Servicebio) and anti-CD163 (Servicebio) overnight at 4°C, followed by incubation with secondary Cy3-labelled antibody (Servicebio) and HRP-labelled antibody (Servicebio) for 50 min at room temperature. Nuclei were counterstained with DAPI.