ELAVL1 Protein Expression Analysis

HCC cells were subjected to Western blot analyses [37 (link)]. Briefly, the lysates were separated by 8% SDS-PAGE gels (Bio-Rad Laboratories, Hercules, CA, USA) and were then transferred to a polyvinylidene difluoride membrane (Bio-Rad Laboratories). The membranes were blotted with primary antibodies against ELAVL1 (Cell Signaling Technology, Danvers, MA, USA), tubulin (Oncogene Science, Cambridge, MA, USA), and Flag (DYKDDDDK) tag (Wako Pure Chemical Industries, Osaka, Japan) and horseradish peroxidase (HRP)-conjugated secondary antibody (GE Healthcare Life Sciences, Chicago, IL, USA). The membranes were developed using Immobilon Western Chemiluminescent HRP Substrate (EMD Millipore, Burlington, MA, USA), and the signals were detected using ChemiDoc XRS Systems (Bio-Rad Laboratories). Band intensity was quantified using Image Lab 4.1 software (Bio-Rad Laboratories).