Figure 5. Effects of mifepristone on adiponectin secretion and expression levels in differentiated adipocytes.

Shown in the upper half of panel A are the results of the Western blot analysis of the culture medium from differentiated adipocytes stimulated with mifepristone. 3T3-L1 adipocytes were serum starved and treated with 1-µM mifepristone for the times indicated. Equal volumes of culture medium were collected and subjected to immunoblot analysis using an antibody specific to adiponectin. Lower half panel A shows the results of densitometric analysis, plotting the fold increase in the degree of adiponectin secretion levels at the time points after mifepristone addition as indicated. Bands corresponding to adiponectin were quantified by densitometry and were normalized to the signals obtained in the absence of mifepristone (open circle, time = 3 day). Panel B shows the results of qRT-PCR assay. Cells were treated with increasing concentrations of mifepristone for 3 days. RNA was then isolated and qRT-PCR was performed. The expression levels of adiponectin and IRS-1 mRNA were normalized to 18S. Shown are the results derived from pooled data, plotting the fold increase in the degree of adiponectin mRNA expression, relative to the values obtained in the absence of mifepristone. Shown are the mean ± S.E.M. of pooled data, derived from five independent in each group. n.s. no significant (p > 0.05); * p < 0.05, ** p < 0.01 versus day 1 in each group; † p < 0.05 versus mifepristone treated cells.