Immunoblotting of HCT-116 and LOVO Cell Lysates

Immunoblotting was performed using standard procedures as previously described [66 (link)]. 30 μg of HCT-116 and LOVO cell lysates were separated by SDS– PAGE using 4–20% Criterion TGX polyacrylamide gels (Bio-Rad, Hercules, CA, USA) and blotted onto a nitrocellulose membrane (Bio-Rad). All primary and secondary antibodies were from Cell Signaling Technology, Danvers, MA, USA. Proteins were detected using the ECL, the ChemiDoc-It2 Imaging System and the VisionWorksLS Software for the analysis (UVP, LLC, Upland, CA, USA). Treatments with AHCC and ETAS were performed for 48 h. Representative images of Western blotting were shown.