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. 2009 Dec 22;59(3):588–599. doi: 10.2337/db09-0796

FIG. 7.

FIG. 7.

Recovery of insulin (Ins) signaling and the inhibition of PEPCK and G6Pase in primary hepatocytes from resveratrol (Resv)-treated IRS2−/− mice. Primary hepatocytes obtained from mice of each condition (treated or not with resveratrol) were cultured as described in research design and methods. A: Cells were serum starved for 4–6 h and further stimulated with 10 nmol/l insulin for 5 min. After cell lysates were prepared, total protein (50 μg) was used for Western blot analysis with the corresponding antibodies against phospho-Akt (Ser 473), phospho-Foxo (Ser 256), total Akt, and PTP1B. A representative experiment corresponding to primary hepatocytes isolated from one animal is shown from three independent experiments performed in triplicate. B: Cells were cultured in serum-free medium for 4–6 h and further stimulated with dex/cAMP (0.5 mmol/l dibutyril cAMP plus 1 μmol/l dexamethasone) in the absence or presence of insulin (10 nmol/l) for 6 h. At the end of the culture time, RNA was isolated and submitted to Northern blot analysis. A representative experiment corresponding to primary hepatocytes isolated from one mouse is shown. The autoradiograms corresponding to three independent experiments performed in hepatocytes were quantitated by scanning densitometry. The value of dex/cAMP-treated cells was set to 100%. Results are expressed as percentage of decrease by insulin of PEPCK and G6Pase mRNAs and are means ± SEM. *P < 0.05, treated vs. untreated. C: Primary hepatocytes obtained from IRS2−/− mice were cultured as described in research design and methods. Cells were treated with 50 μmol/l resveratrol for 24 h, serum-starved for 2 h, and then stimulated with 10 nmol/l insulin for 5 min. After cell lysates were prepared, total protein (50 μg) was used for Western blot analysis with the corresponding antibodies against phospho-Akt (Ser 473), phospho-Foxo (Ser 256), total Akt, and PTP1B. A representative experiment corresponding to primary hepatocytes isolated from one animal is shown from three independent experiments performed in triplicate.