4.5. Western Blot Analysis

The AML12 cells and liver specimens obtained from the mice were lysed using the EzRIPA Lysis kit (ATTO Corporation; Tokyo, Japan), and quantified by Bradford reagent (Bio-Rad, Hercules, CA, USA). Proteins were visualized by western analysis using the following primary antibodies (1:1000 dilution) from Cell Signaling Technology (Beverly, MA, USA) and then with HRP-conjugated secondary antibodies (1:2000 dilution) from Vector laboratories (Burlingame, CA, USA). Specific immune complexes were detected using the Western Blotting Plus Chemiluminescence Reagent (Millipore, Bedford, MA, USA).

Primary antibodies against proliferating cell nuclear antigen (PCNA), phosphorylated-signal transducer and activator of transcription 3 (p-STAT3), STAT3, hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), B-cell leukemia-extra large (Bcl-xL), dynamin related protein 1 (DRP-1), and β-actin were all obtained from Cell Signaling Technology (Beverly, MA, USA), and Opa1 mitochondrial dynamin like GTPase (OPA-1) was obtained from Santa Cruz biotechnology (Santa Cruz, CA, USA). Horseradish peroxidase (HRP)-conjugated secondary antibody were obtained from Cell Signaling Technology (Beverly, MA, USA).