Western Blot Analysis of Key Cellular Proteins

Protein extraction was performed based on our previous report [14 (link)]. Primary antibodies were diluted 1:2000 and incubated overnight at 4 °C. Secondary antibodies diluted 1:5000 were added and incubated at room temperature for 1 h. Signals were detected using ECL detection reagent (Millipore Corporation, Burlington, MA, USA) following the manufacturer’s instructions. The primary antibodies used were as follows: mouse anti-β-catenin, mouse anti-lamin A/C (BD Biosciences, San Jose, CA, USA), mouse anti-c-myc, mouse anti-cyclin D1, rabbit anti-HDAC1, rabbit anti-HIF-1α and rabbit anti-β-actin (GeneTex, Irvine, CA, USA). Horseradish peroxidase-conjugated rabbit anti-mouse or goat anti-rabbit secondary antibodies (GeneTex) were used as appropriate. Detailed information is given in the Supplementary Materials.