Anti pcna sc 56

Tumor tissues were stained with H&E as per standard protocols and analyzed under a light microscope (Olympus). For all staining protocols, the tissue sections were deparaffinized, rehydrated, and washed in 1% PBS–Tween 20. For IHC, the sections were soaked into 3% hydrogen peroxide to block endogenous peroxidases, blocked with 5% goat serum, and incubated with the primary antibodies overnight at 4°C. The sections were then incubated with streptavidin-HRP for 1 hour, stained with diaminobenzidine substrate, and counterstained with hematoxylin. For IF, the slides were stained with primary antibodies overnight at 4^°C, incubated with fluorescently labeled secondary antibody for 1 hour the next day, and then counterstained with DAPI for 5 min. The TUNEL assay was performed according to the manufacturer’s protocol using a TMR (red) TUNEL Cell Apoptosis or FITC TUNEL Cell Apoptosis Detection Kit (G1501-50, G1502-50; Servicebio Technology, Wuhan, China). Images were acquired by fluorescence microscopy (Olympus). The antibodies used here included anti-PCNA (sc-56; Santa Cruz, CA, USA), anti-CD4 (sc-13573; Santa Cruz), anti-CD8 (sc-18913; Santa Cruz), anti-granzyme B (sc-8022; Santa Cruz), and anti-IFN-γ (15365-1-AP; Proteintech, USA). The secondary antibodies used in IF were purchased from Thermo Fisher Scientific (A11037, A21135, and A21209).