FIGURE 5.

AMPK mediates tadalafil inhibition of high glucose-stimulated mTOR complex 1 activation. Podocytes were preincubated with Compound C (CC) for 30 min. Cells were incubated with 5 or 30 mm glucose (GLU) for 5 (A–C) or 30 min (D) with or without preincubation with tadalafil (TAD) for 8 h. Cell lysate proteins were immunoblotted with specific antibodies. A, antibody against phospo-4E-BP1 (Thr-37/46) (P-4E-BP1) or 4E-BP1. B, antibody against phospho-p70S6 kinase (Thr-389) (P-S6K) and p70S6 kinase (S6K). C, antibody against phosphorylated eEF2 kinase (Ser-366) (P-eEF2K) and eEF2 kinase (eEF2K). D, antibody against phospho-eEF2 (Thr-56) (P-eEF2) or eEF2. In A–D, composite data from three to five experiments are shown in histograms; error bars represent S.E.