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Fig. 9 | Molecular Medicine

Fig. 9

From: Tissue Distribution and Regulation of Plasminogen Activator Inhibitor-1 in Obese Mice

Fig. 9

Effect of insulin on PAI-1 expression by cultured 3T3-L1 adipocytes

3T3-L1 adipocytes were grown and differentiated in 100-mm tissue culture plates as described (36). Total RNA was isolated from untreated cells (), and cells treated with 100 nM insulin () for the indicated times, and the steady-state levels of PAI-1 mRNA were determined using quantitative PCR (n = 6 ± SD). Inset: Conditioned medium (20 µl) from untreated cells and cells treated with insulin for various times were electrophoresed under non-reducing conditions on a 9% SDS-PAGE gel and transferred to nitrocellulose membranes. The membranes were analyzed for PAI-1 by Western blotting using a polyclonal rabbit anti-mouse PAI-1 antiserum and the enhanced chemiluminescence detection system. mPAI-1 refers to the recombinant murine PAI-1 antigen used as a standard.

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