Western blotting
PBMCs were stimulated with various cytokines for indicated time. The
extracted protein was subjected to 10% SDS-PAGE and transferred onto
PVDF membranes (Millipore). The following primary antibodies were used:
rabbit anti-phosphorylated STAT1 (Cell Signaling Technology) and rabbit
anti-STAT1 (Cell Signaling Technology), rabbit anti–phosphorylated
STAT3 (Cell Signaling Technology), mouse anti-STAT3 (BD), rabbit
anti-TYK2(Abcam, cat223733), and that against β-actin rabbit mAb
horseradish peroxidase (HRP)-conjugated (Cell Signaling Technology). The
HRP-conjugated goat anti-rabbit IgG secondary antibody (Cell Signaling
Technology) and the HRP-conjugated goat anti-mouse IgG secondary
antibody (ZSGB-BIO, China).