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. 2016 Jul 12;291(35):18536–18546. doi: 10.1074/jbc.M116.726836

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© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 6. — ATF4 inhibits AMPK activity and promotes TG accumulation via increasing TRB3 expression in vitro under ethanol treatment. A–E, AML-12 cells were infected with Ad-scrambled (−Ad-shTRB3) or Ad-shTRB3 (+Ad-shTRB3), Ad-GFP (−Ad-DN ATF4) or Ad-DN ATF4 (+Ad-DN ATF4), or Ad-GFP (−Ad-TRB3) or Ad-TRB3 (+Ad-TRB3) as indicated for 48 h, followed without (−EtOH) or with 100 mm ethanol treatment (+EtOH) for 24 h. Means ± S.E. (error bars) shown are representative of at least three independent in vitro experiments. Statistical significance was determined by Student's t test for the effect of Ad-shTRB3 or Ad-DN ATF4 versus corresponding control under the same treatment (with or without ethanol) (*, p < 0.05); with versus without ethanol treatment in the presence of the same adenovirus infection in A–D or with versus without Ad-TRB3 in the presence of Ad-DN ATF4 in E and F (#, p < 0.05). A, C, and E, Atf4 and/or Trb3 mRNAs and TRB3, p-AMPK, AMPK, p-ACC, ACC, and ATF4 proteins (left, Western blotting; right, quantitative measurements of TRB3, p-AMPK, and p-ACC relative to their total proteins or actin); B, D, and F, TG contents.