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. Author manuscript; available in PMC: 2014 Sep 1.
Published in final edited form as: Circ Heart Fail. 2013 Jul 19;6(5):10.1161/CIRCHEARTFAILURE.113.000338. doi: 10.1161/CIRCHEARTFAILURE.113.000338

Figure 1. Cardiomyocyte-restricted Csn8 knockout initiated in adult mice impairs deneddylation activities in the heart.

Figure 1

At 5 days after the first tamoxifen injection, ventricular myocardium was sampled from Csn8flox/flox (CTL) or Csn8flox/flox::MerCreMertg (CSN8CKO) littermate mice for western blot analyses. Representative images are shown with each lane representing a mouse. α-Tubulin or GAPDH was probed for loading control. (A) Changes in CSN8, CSN1 and CSN5. AU: arbitrary unit. (B) Increases in the neddylated form (arrow) and decreases in the native form (*) of cullin1 (Cul1), Cul 2, Cul3, and Cul4A in CSN8CKO hearts. To verify the identity of the neddylated cullins, cultured neonatal rat cardiomyocytes were treated with a specific NEDD8 E1 inhibitor MLN4924 (MLN, 1µM) (Active Biochem, Maplewood, NJ) or vehicle (−) for 24 hours. MLN4924, which inhibits neddylation, abolished the slow-migrating band of all cullins, confirming their identity as neddylated cullins. (C) Increased Nedd8 conjugates in CSN8CKO hearts.