Journal: PLoS ONE

Article Title: The Ginsenoside 20-O-β-D-Glucopyranosyl-20(S)-Protopanaxadiol Induces Autophagy and Apoptosis in Human Melanoma via AMPK/JNK Phosphorylation

doi: 10.1371/journal.pone.0104305

Figure Lengend Snippet: (A) Phosphorylation of AMPKα is increased by GPD in a dose-dependent manner. (B) GPD upregulates phosphorylated JNK1/2 in a dose-dependent manner. (C) Levels of phosphorylated c-Jun, a well-known downstream of JNK, peak at 24 h after GPD treatment. (D) The phosphorylation of c-Jun after GPD treatment was dramatically decreased by co-treatment with the JNK inhibitor SP600125, whereas it decreased slightly in the presence of SB202180, a p38 inhibitor. (E) Activated AMPKα negatively regulated the phosphorylation of mTOR in a time-dependent manner. Cells (5×10 5 /100 mm dish) were incubated with various concentrations of GPD for the indicated durations. (F) GPD induces the phosphorylation of Bcl-2 and the dissociation of Beclin-1 from Bcl-2. Western blot analysis was performed using specific antibodies as described in . β-actin was used to verify equal protein leading.

Article Snippet: The antibody against LC3 was purchased from ABGENT Inc. (San Diego, CA), while antibodies against phosphorylated AMPKα (Thr172), total AMPKα, phosphorylated c-Jun NH2-terminal kinases (JNKs; Thr183/Tyr185), total JNK, Beclin-1, phosphorylated Bcl-2, cytochrome C, PARP, caspase-3, 7, 9, phosphorylated ATF2, ATF, phosphorylated c-Jun (Ser73), total c-Jun, phosphorylated mTOR (Ser2448) and total mTOR were purchased from Cell Signaling Technology (Danvers, MA).

Techniques: Incubation, Western Blot