Figure 10.

Inhibition of binding of cytoplasmic factor to TGF-β1 5′-UTR by wortmannin or rapamycin. Radiolabeled probe generated against sequences in the 5′-UTR of TGF-β1 mRNA spanning +1 to +147 was incubated with cytoplasmic extract of HK-2 cells treated with insulin alone (5 μg/ml) or insulin in the presence of either wortmannin (10 nmol/L) or rapamycin (30 nmol/L) for 48 hours. The mixture was resolved on a nondenaturing polyacrylamide gel. Competition was with an excess of unlabeled RNA probe.