c-Jun N-terminal kinase-mediated Rubicon expression enhances hepatocyte lipoapoptosis and promotes hepatocyte ballooning

Figure 4 Palmitate induces expression of both p62 and Rubicon, and PA-induced Rubicon expression is mediated by c-Jun N-terminal kinase phosphorylation. A: Whole cell lysates were prepared from AML12 cells treated with PA (400 or 800 μmol/L) for 4 h in the presence or absence of SP600125. Immunoblotting analyses were performed for p62, LC3, and Rubicon. β-actin was used as the loading control; B: AML12 cells were incubated with 800 μmol/L PA for 4 h in the presence or absence of 30 μmol/L SP600125. p62 antibody was used as the primary antibody, and Alexa Fluor^® 488-labeled secondary antibody was used for detecting p62 antibody. Samples were visualized using an EVOS microscope; C: Total RNA was prepared from AML12 cells treated with PA (800 μmol/L) for 4 h in the presence or absence of 30 μmol/L SP600125. Vehicle-treated cells were used as the control (CT). Both p62 and Rubicon mRNA were quantified by RT-qPCR, normalized to Actin, and expressed as the fold-change vs control cells without SP600125. All of the above experiments were repeated three times and representative results are shown. The quantitative data are presented as the mean ± SD; ^aP < 0.05 vs control.