Skip to main content
. Author manuscript; available in PMC: 2015 May 3.
Published in final edited form as: Endocrinology. 2005 Oct 6;147(1):141–154. doi: 10.1210/en.2004-1649

Fig. 6.

Fig. 6

Inhibition of adipogenic differentiation in 3T3-L1 cells by testosterone is blocked by overexpression of TCF4 or a dominant negative TCF4 construct. 3T3-L1 cells were transfected either with full-length TCF4 or a dominant negative TCF4 (Dn-TCF4) cDNA construct or a control vector and were allowed to differentiate either in the presence (+T) or in the absence (−T) of testosterone (100 nm) for 12 d. Cells were fixed and stained with Oil Red O and quantitative image analysis was performed. A, Oil Red O staining: Representative photographs of Oil Red O-stained adipocytes are shown from different treatment groups (panel A). B, The graph shows the quantitative image analysis data obtained after averaging 20 fields from each treatment group, mock (Cont), TCF4 transfection (TCF4), and Dn-TCF4 transfection (Dn-TCF4) (P values vs. control: *, P < 0.05; IOD denotes integrated optical density; T denotes testosterone). C, Real-time quantitative RT-PCR analysis demonstrating the effects of testosterone on C/EBP-α and AP2 mRNA expression in TCF4 and Dn-TCF4 overexpressed 3T3-L1 cells 3T3-L1 cells were transfected with full-length TCF4 or Dn-TCF4 or a control vector plasmid and were allowed to differentiate in adipogenic medium (3 d) then GM with or without testosterone (100 nm) for 9 d as in panel A. Total cellular RNA was isolated and C/EBP-α and AP2 mRNA expression was analyzed by real-time RT-PCR. [*, P < 0.03 and **, P < 0.0005 denotes P values vs. control group (no T) in the upper panel; whereas #, P < 0.05, ##, P < 0.01 and ###, P < 0.002 denotes P values vs. control group (no T) in the lower panel.]