Anti interferon γ ifn γ

PC-3 cells were cultured in the cell culture chamber. The cells were fixed in 4% paraformaldehyde for 15 min, permeabilized using 0.2% triton X-100 for 10 min, followed by staining with rabbit anti-human γH2AX, rabbit anti-human 53BP1, mouse anti-human phospho-histone H3, rabbit anti-human phospho-p65 (Cell signal technology) and mouse anti-human PD-L1 antibody (Abcam) or normal control IgG 6–8 hours at 4°C. Then, Alexa Flour 488-conjugated goat anti-rabbit IgG and Alexa Flour 594-conjugated goat anti-mouse IgG were added and incubated for 1 hour at 37°C. The nucleus was stained with 4'6-diamidino-2-phenylindole and mounted with ProLong Gold Antifade Reagent (Life Technologies). The images were analyzed using a Zeiss LSM 880 Confocal Imaging System (Zeiss, Jena, Germany).31 (link)
Paraffin-embedded tissues were sectioned and subjected to immunostaining using primary antibodies such as anti-PD-L1, anti-CD45, anti-phospho-ATM, anti-phospho-p65, anti-CD3 and anti-interferon-γ (IFNγ) (Abcam). The isotype-matched primary antibodies served as the controls. Antibody binding was visualized using fluorescence-labeled secondary antibodies under a Zeiss LSM 880 Confocal Imaging System.