Figure 5.

SIRT1 positively regulates AMPK activation in cultured skin keratinocytes. HaCaT cells were pre-treated with sirtinol (2 mM) or nicotinamide (Nico, 10 mM) for 1 hr, followed by 20 mJ/cm² of UV for indicated time, p-AMPK (Thr 172) and total-AMPK activation were detected by Western blot (A and B). HaCaT cells were pre-treated with sirtinol (2 mM) or nicotinamide (Nico, 10 mM) for 1 hr, followed by 20 mJ/cm² of UV and incubated for 0.5 and 2.0 hrs, p-ACC (Ser 79), p-PFK-2 (Ser 466) and β-actin were detected by Western blot (C), p-ACC was quantified in (D). HaCaT cells were treated with nicotinamide (Nico, 10 mM) or AMPK inhibitor Compound C (AMPKi, 10 μM) for 0.5, 2.0 hrs, followed by resveratrol (Rev, 10 μM) treatment for 0.5, 1.0 and 2.0 hrs, p-AMPK (Thr 172), p-ACC (Ser 79) and T-AMPK were detected by Western blot (E and F). The data in figures represent mean ± S.E. of three independent experiments. The symbol ‘^#’ means P < 0.05 with UV- or H₂O₂-treated group.