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. Author manuscript; available in PMC: 2013 Oct 3.
Published in final edited form as: Cell Metab. 2012 Oct 3;16(4):462–472. doi: 10.1016/j.cmet.2012.08.011

Figure 4. Interference with PPARγ causes decreased Cullin-3 and Nedd8-Cullin-3.

Figure 4

(A–B) Protein levels of Cullin-3 and neddylated Cullin-3, Cullin-1 and neddylated Cullin-1, and total Nedd8-conjugates (Nedd8) in medial aortic tissue from S-P467L and NT mice determined by Western blot (n=8–9) and their quantification. Neddylation is the covalent attachment of Nedd8 onto a target protein. The lower molecular weight band of the doublet (~85 kDa) is unneddylated Cullin-3 or Cullin-1 (denoted as Cul3 or Cul1) whereas the higher band (~95 kDa) is the neddylated Cullin-3 or Cullin-1 product (denoted as Nedd8-Cul3 or Nedd8-Cul1). Cullin-3 mRNA expression determined by qPCR (n=10). See also Table S1. * p<0.05 S-P467L vs. NT. Error bars represent SEM. (C–D) Primary rat aortic smooth muscle cells treated with the Nedd8-activating enzyme inhibitor MLN4924 (1 μmol/L) or DMSO over a time-course of 24 hr. Protein lysates were assessed for Cullin-3, Cullin-1, Nedd8 and RhoA by Western blot and quantified (n=4). U, untreated; V, vehicle (DMSO, 24 hr). “NS” indicates non-specific band. * p<0.05 MLN4924-treated vs. vehicle-treated. Error bars represent SEM.