Effects of SHT on the expression levels of melanogenic proteins, p-PKA, PKA, p-CREB, CREB, p-MAPKs, and MAPKs after stimulation with α-MSH in B16F10 cells. (A) B16F10 cells were pre-incubated with or without 250 and 500 μg/ml SHT for 12 h, and then stimulated with 1 μM of α-MSH for 36 h. After harvest of cells, lysates were prepared and subjected to Western blot analysis with the anti-tyrosinase, anti-TRP-1, anti-TRP-2, or anti-MITF antibody. (B-C) Cells were pre-incubated with or without 500 μg/ml SHT for 12 h, and then further stimulated with 1 μM of α-MSH for 15, 30, and 60 min. Changes in the level of p-PKA and p-CREB (B) and in the level of p-MAPKs including p-p38, p-ERK, and p-JNK (C) were examined by Western blot analysis followed by quantitation with ImageJ. Data are representative of two independent experiments.