Overexpression of Ccdc80 impairs adipogenesis. 3T3-L1 cells were
infected with adenovirus encoding either LacZ (Ad-LacZ, white
bars) or mouse Ccdc80 (Ad-Ccdc80, black bar) at various
multiplicity of infection (m.o.i.). A, expression of Ccdc80 was
determined by real-time PCR. B, secretion of Ccdc80. Conditioned
medium from postconfluent (Day 0) and fully differentiated (Day 8) 3T3-L1
infected with adenovirus at a m.o.i. of 2000 was analyzed by Western blotting
using an antibody raised against amino acids 840-854 of Ccdc80.
Arrows indicate the full-length (∼140 kDa) and cleaved fragments
(∼50 and ∼22 kDa) of Ccdc80 in conditioned medium from postconfluent
(Day 0) and fully differentiated (Day 8) adipocytes.
C, lipid accumulation. D, expression of adipogenic markers.
Expression of aP2, C/EBPα, and PPARγ was determined by real-time
PCR. E, lipid accumulation in 3T3-L1 cell lines expressing a NS shRNA
or Ccdc80 shRNA infected with adenovirus encoding either LacZ or Ccdc80.
F, effect of TZD on adipogenesis. Postconfluent (Day 0) 3T3-L1 cells
infected with adenovirus at an m.o.i. of 2000 were differentiated with
adipogenic inducers (dexamethasone, IBMX, and insulin) in the presence or
absence of rosiglitazone (100 nm). C, E, and
F, lipid accumulation was visualized by oil red O staining.
Macroscopic and microscopic (100× magnification) images are shown.
*, p < 0.05 versus Ad-LacZ.