Setdb1 antibody

Tissues were fixed overnight in 4% paraformaldehyde and embedded in paraffin. All IHC staining was performed on 4-µm sections. After deparaffinization, rehydration, antigen unmasking, and endogenous peroxidase blocking, sections were blocked in 5% BSA with 0.1% Triton X-100 in PBS. Sections were incubated overnight at 4°C with primary antibodies. The details of antibodies are as follows: CD3 antibody (Abcam; cat #ab5690; 1:200); CD4 antibody (Abcam; cat #ab183685; 1:200); CD8 antibody (Cell Signaling Technology; cat #98941; 1:200); Foxp3 antibody (Cell Signaling Technology; cat #12653; 1:200); SETDB1 antibody (Abcam; cat #ab12317; 1:200); and FOXM1 antibody (Abcam; cat #ab232649; 1:200). Tissue sections were then stained using an Immunohistochemistry kit (Servicebio; cat #G1215; cat #G1215-200T; cat #G1216-200T). Tissues were observed under a microscope and photographed after being counterstained with hematoxylin, dried, and mounted. For assessment of CD3⁺, CD4⁺, CD8⁺, and Foxp3⁺ T cell infiltration, positively stained cells were counted from five high-power random fields of each sample, and the numbers averaged for each field. The IHC scores for SETDB1 and FOXM1 staining are assigned using a semi-quantitative five-category grading system as previously described (Sun et al., 2018 (link)).