Immunoblotting Analysis of Protein Lysates

The primary tissue lysates and whole cell lysates were prepared for immunoblotting as described previously 25 (link). Equal amounts of total proteins (30 µg) were subjected to SDS-PAGE and immunoblotting analysis with specific antibodies. The primary antibody against Cyclin D1 was purchased from Cell Signaling Technology (Danvers, MA). Anti-USP5 antibody was obtained from Proteintech Group (Wuhan, China). Anti-TUFM and anti-EBF1 antibodies were purchased from ABclonal Biotechnology (Hubei, China). Anti-Flag, Myc and HA antibodies were purchased from Medical & Biological Laboratories (Tokyo, Japan). Anti-GAPDH antibody was purchased from Abgent Biotechnology (Suzhou, China). Horseradish peroxidase-conjugated anti-mouse and anti-rabbit IgG antibodies were purchased from Beyotime Biotechnology (Nantong, China).