Western Blotting of N-Myc and β-Actin

Western blotting was performed as previously described [19 (link)]. Briefly, 60 µg of total protein was separated on 10% polyacrylamide/SDS gel and transferred to PVDF membranes, and the membranes were blocked with 5% (w/v) bovine serum albumin (BSA) in tris buffered saline (TBS) (+0.1% Tween-20). The primary antibodies included N-Myc (Cell Signaling Technology Cat # 84406, RRID: AB_2800038, Danvers, MA, USA), β-Actin (Cell Signaling Technology Cat # 4970, RRID: AB_2223172), and horseradish peroxidase-conjugated secondary antibody: anti-rabbit IgG (Cell Signaling Technology Cat# 7074, RRID: AB_2099233).