Anti cd8a

Patient tissue samples and mouse tumor tissues were fixed in neutral buffered formalin (10%), paraffin-embedded, and sectioned at 4 μm thickness. For IHC, the samples were incubated with the following primary antibodies at 4 °C overnight: anti-USP8 (27791-1-AP, Proteintech), anti-PD-L1 (66248-1-Ig, Proteintech), anti-CD8a (98941, Cell Signaling Technology), anti-Granzyme B (44153 S, Cell Signaling Technology), anti-cleaved caspase-3 (9661, Cell Signaling Technology), and anti-Ki67 (12202, Cell Signaling Technology). The next day, the sections were incubated with secondary antibodies conjugated to horseradish peroxidase (HRP). The samples were visualized using a diaminobenzidine (DAB) chromogen kit (BDB2004, Biocare, Pacheco, CA, USA). Image Scope software (Leica Biosystems, Wetzlar, Germany) was used to capture representative images of the tumors. The quantitative results for IHC staining were obtained using ImageJ software and GraphPad Prism 8 (GraphPad Software, San Diego, CA, USA). Microarray slides of PDAC tissue were immunostained using antibodies against USP8 (27791-1-AP, Proteintech) and CD274 (ab205921, Abcam). 3DHISTECH QuantCenter 2.1 software (3Dhistech, Budapest, Hungary) was used to quantify the IHC results.