Western blotting.
Whole-cell lysates of patient PBMCs after anti–PD-1 treatment or PBMCs from healthy donors were separated by SDS-polyacrylamide electrophoresis and then probed with antibodies against p-PKCθ (BD Biosciences, 612735), PKCθ (Cell Signaling Technology, 13643), and GAPDH (ZSGB-BIO, TA-08).