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Figure 2


Fig. 2. Representative time-course gene expression profiles in 3T3-L1 cells. Differentiated (DIFF) cells incubated for 1, 3 or 6 days with or without (diamonds and black line) 10 ng/ml TNF{alpha} (squares and short dashed line) or 0.5 µM FCCP (triangle and long dashed line). The gene expression profile as a function of time determined by cDNA microarray is illustrated for (A) AGT, (B) CHOP-10, (C) CPT 2 and (D) PPAR{gamma}. Results are expressed as relative transcript abundance, transformed to log2. Set of genes displaying profiles similar to AGT and CTP 2 are listed between A and B and C and D, respectively. (E,F) Effect of FCCP or TNF{alpha} on the DNA-binding activity of PPAR{gamma} and C/EBP{alpha} in adipocytes. DNA-binding activity for (E) PPAR{gamma} and (F) C/EBP{alpha} was determined in nuclear proteins prepared from differentiated (DIFF) and undifferentiated cells (CTL) and then incubated for an extra 24 (white columns) or 72 hours (black columns) with or without 10 ng/ml TNF{alpha}, 0.5 µM FCCP or 10 µM isoproterenol using a Trans-AM assay. Results are expressed as percentages of DNA-binding activity found in differentiated cells as means ± s.d. for 3 experiments. *P<0.5, **P<0.01 and ***P<0.001, significantly different from DIFF cells. (G) Western blot analysis for PPAR{gamma} was performed using the same conditions as above. Equal protein loading was controlled by the TBP immunodetection.





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