Anti akt ab

Cells were cultured in 10-cm cell culture dishes and treated with aloperine. DMSO was used as a negative control. The whole cellular extract was subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis, and the separated proteins were electrically transferred to a PVDF membrane (Millipore Corporation, USA). The membrane was blocked with primary antibodies (anti-LC3 Ab; Abcam, USA; anti-GAPDH Ab; GeneTex, USA; anti-AMPK α-1 Ab; Cell signaling, USA; anti-phosphorylated-AMPK α-1 (Thr 172) Ab; Cell Signaling, USA; anti-Akt Ab; Santa Cruz, USA; anti-phosphorylated-Akt (Ser 473) Ab; Santa Cruz, USA; anti-mTOR Ab; Cell Signaling, USA; anti-phosphorylated-mTOR (Ser 2448) Ab; Cell Signaling, USA; anti-p70S6K Ab; Cell Signaling, USA; anti-phosphorylated-p70S6K (Thr 389) Ab; Cell Signaling; anti-p62/SQSTM1 Ab; Abgent, USA; anti-Erk Ab; Cell Signaling, USA; anti-phosphorylated-Erk (Thr 202/204) Ab; Cell Signaling, USA; anti-JNK Ab; Cell Signaling, USA; anti-phosphorylated-JNK (Thr 183/Tyr 185) Ab; Cell Signaling, USA; anti-p38 Ab; Cell Signaling, USA; and anti-phosphorylated (Thr 180/Tyr 182) Ab; Cell Signaling Ab, USA) and was analyzed using the BioSpectrum 800 Imaging System (UVP, CA, USA).

Anti‐GAPDH Ab was purchased from Santa Cruz Biotechnology. Anti‐Akt Ab, anti‐p‐Akt (Thr308) Ab, anti‐PI3‐kinase p85 Ab, anti‐mouse‐HRP secondary Ab, anti‐His antibody anti‐SLC1A5 antibody, anti‐goat‐HRP secondary Ab, anti‐ATF4 antibody, and anti‐rabbit‐HRP secondary Ab were purchased from Cell Signaling Technology. Anti‐COL1A1 antibody and anti‐alpha‐SMA antibody were purchased from Abcam. Biotinylated Concanavalin A (Con A) and Biotinylated PHA‐L (Phaseolus Vulgaris Leucoagglutinin) were purchased from Vector Laboratories.