Anti tyrp2

Whole-cell extracts were prepared using a radioimmunoprecipitation assay lysis buffer (Thermo Fisher Scientific, USA) containing a protease inhibitor cocktail (Roche Diagnostics, USA). Extracts were subjected to SDS-PAGE and subsequently transferred to a nitrocellulose membrane. The membrane was then incubated with 5 % skim milk for 1 h, followed by incubation with each indicated primary antibody at 4 °C. For protein detection, horseradish peroxidase-conjugated secondary antibodies (Cell Signaling Technology, USA) were used, followed by enhanced chemiluminescence (ECL, Pierce, Thermo Fisher Scientific) and autoradiography. β-actin was used as an endogenous internal control. The primary antibodies used in these experiments were as follows. Anti-Mitf, anti-tyrosinase, anti-TYRP1, anti-TYRP2, and anti-p38 antibodies were purchased from Santa Cruz Biotechnology (USA). Anti-ERK, anti-p-ERK (Thr202/Tyr204), anti-p-p38 (Thr180/Tyr182), anti-JNK, anti-p-JNK (Thr183/Tyr185) and anti-p-PKA (Thr198) antibodies were purchased from Cell Signaling Technology. Anti-PKA antibody was purchased from Abcam (UK).