TFEB translocates to the nucleus upon AMPK stimulation independently of mTOR in mouse primary hepatocytes. A) AMPKα1α2+/+ or AMPKα1α2−/− mouse primary hepatocytes were isolated and cells were treated with 10 µM 991, 300 µM AICAR, 30 µM C13, or rapamycin 0.05 µM for 1 or 4 h. Western blot analysis of the indicated proteins was performed on cell lysates. B) Primary hepatocytes were isolated from AMPKα1α2−/− mice and were untreated (No) or coinfected with 3 adenoviruses (1:3 multiplicity of infection) in order to rescue the expression of the 3 subunits of AMPK (Flag-α2, β1, and γ1). Sixteen hours postinfection, the primary hepatocytes were treated for 1 h with vehicle (DMSO) or 10 µM 991. Cell lysates (20 µg) were used for Western blot analysis with the indicated antibodies. Images are representative of n = 3.
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