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( A ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice under chow diet. Expression was assessed by qRT-PCR ( n = 9–11). Mice were fed a chow diet and were sacrificed 8 weeks after infection with AAV. ( B ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice after 10 weeks on HFD. Expression was assessed by qRT-PCR ( n = 7). Mice were fed an HFD for 2 weeks before infection with AAV and were sacrificed 8 weeks later. ( C ) Protein levels of SREBP-1c, SCD1, and CHAC1 in livers of mice under HFD. ( D ) Relative expression levels of Chac1 -regulated genes in primary hepatocytes under 100 μM free fatty acid (FA) treatment. Expression was assessed by qRT-PCR ( n = 3). Adenovirus (Adv): 400 μL/mL medium. Cells were harvested 3 days postinfection. ( E ) Protein levels of <t>SPP1,</t> SCD1, and CHAC1 in primary hepatocytes under 100 μM free FA treatment. Primary hepatocytes were infected with Adv overexpressing GFP or Chac1 . Adv: 400 μL/mL medium. Cells were harvested 3 days postinfection. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
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( A ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice under chow diet. Expression was assessed by qRT-PCR ( n = 9–11). Mice were fed a chow diet and were sacrificed 8 weeks after infection with AAV. ( B ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice after 10 weeks on HFD. Expression was assessed by qRT-PCR ( n = 7). Mice were fed an HFD for 2 weeks before infection with AAV and were sacrificed 8 weeks later. ( C ) Protein levels of SREBP-1c, SCD1, and CHAC1 in livers of mice under HFD. ( D ) Relative expression levels of Chac1 -regulated genes in primary hepatocytes under 100 μM free fatty acid (FA) treatment. Expression was assessed by qRT-PCR ( n = 3). Adenovirus (Adv): 400 μL/mL medium. Cells were harvested 3 days postinfection. ( E ) Protein levels of <t>SPP1,</t> SCD1, and CHAC1 in primary hepatocytes under 100 μM free FA treatment. Primary hepatocytes were infected with Adv overexpressing GFP or Chac1 . Adv: 400 μL/mL medium. Cells were harvested 3 days postinfection. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
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( A ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice under chow diet. Expression was assessed by qRT-PCR ( n = 9–11). Mice were fed a chow diet and were sacrificed 8 weeks after infection with AAV. ( B ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice after 10 weeks on HFD. Expression was assessed by qRT-PCR ( n = 7). Mice were fed an HFD for 2 weeks before infection with AAV and were sacrificed 8 weeks later. ( C ) Protein levels of SREBP-1c, SCD1, and CHAC1 in livers of mice under HFD. ( D ) Relative expression levels of Chac1 -regulated genes in primary hepatocytes under 100 μM free fatty acid (FA) treatment. Expression was assessed by qRT-PCR ( n = 3). Adenovirus (Adv): 400 μL/mL medium. Cells were harvested 3 days postinfection. ( E ) Protein levels of <t>SPP1,</t> SCD1, and CHAC1 in primary hepatocytes under 100 μM free FA treatment. Primary hepatocytes were infected with Adv overexpressing GFP or Chac1 . Adv: 400 μL/mL medium. Cells were harvested 3 days postinfection. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
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( A ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice under chow diet. Expression was assessed by qRT-PCR ( n = 9–11). Mice were fed a chow diet and were sacrificed 8 weeks after infection with AAV. ( B ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice after 10 weeks on HFD. Expression was assessed by qRT-PCR ( n = 7). Mice were fed an HFD for 2 weeks before infection with AAV and were sacrificed 8 weeks later. ( C ) Protein levels of SREBP-1c, SCD1, and CHAC1 in livers of mice under HFD. ( D ) Relative expression levels of Chac1 -regulated genes in primary hepatocytes under 100 μM free fatty acid (FA) treatment. Expression was assessed by qRT-PCR ( n = 3). Adenovirus (Adv): 400 μL/mL medium. Cells were harvested 3 days postinfection. ( E ) Protein levels of <t>SPP1,</t> SCD1, and CHAC1 in primary hepatocytes under 100 μM free FA treatment. Primary hepatocytes were infected with Adv overexpressing GFP or Chac1 . Adv: 400 μL/mL medium. Cells were harvested 3 days postinfection. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
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( A ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice under chow diet. Expression was assessed by qRT-PCR ( n = 9–11). Mice were fed a chow diet and were sacrificed 8 weeks after infection with AAV. ( B ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice after 10 weeks on HFD. Expression was assessed by qRT-PCR ( n = 7). Mice were fed an HFD for 2 weeks before infection with AAV and were sacrificed 8 weeks later. ( C ) Protein levels of SREBP-1c, SCD1, and CHAC1 in livers of mice under HFD. ( D ) Relative expression levels of Chac1 -regulated genes in primary hepatocytes under 100 μM free fatty acid (FA) treatment. Expression was assessed by qRT-PCR ( n = 3). Adenovirus (Adv): 400 μL/mL medium. Cells were harvested 3 days postinfection. ( E ) Protein levels of <t>SPP1,</t> SCD1, and CHAC1 in primary hepatocytes under 100 μM free FA treatment. Primary hepatocytes were infected with Adv overexpressing GFP or Chac1 . Adv: 400 μL/mL medium. Cells were harvested 3 days postinfection. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
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( A ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice under chow diet. Expression was assessed by qRT-PCR ( n = 9–11). Mice were fed a chow diet and were sacrificed 8 weeks after infection with AAV. ( B ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice after 10 weeks on HFD. Expression was assessed by qRT-PCR ( n = 7). Mice were fed an HFD for 2 weeks before infection with AAV and were sacrificed 8 weeks later. ( C ) Protein levels of SREBP-1c, SCD1, and CHAC1 in livers of mice under HFD. ( D ) Relative expression levels of Chac1 -regulated genes in primary hepatocytes under 100 μM free fatty acid (FA) treatment. Expression was assessed by qRT-PCR ( n = 3). Adenovirus (Adv): 400 μL/mL medium. Cells were harvested 3 days postinfection. ( E ) Protein levels of <t>SPP1,</t> SCD1, and CHAC1 in primary hepatocytes under 100 μM free FA treatment. Primary hepatocytes were infected with Adv overexpressing GFP or Chac1 . Adv: 400 μL/mL medium. Cells were harvested 3 days postinfection. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.
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( A ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice under chow diet. Expression was assessed by qRT-PCR ( n = 9–11). Mice were fed a chow diet and were sacrificed 8 weeks after infection with AAV. ( B ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice after 10 weeks on HFD. Expression was assessed by qRT-PCR ( n = 7). Mice were fed an HFD for 2 weeks before infection with AAV and were sacrificed 8 weeks later. ( C ) Protein levels of SREBP-1c, SCD1, and CHAC1 in livers of mice under HFD. ( D ) Relative expression levels of Chac1 -regulated genes in primary hepatocytes under 100 μM free fatty acid (FA) treatment. Expression was assessed by qRT-PCR ( n = 3). Adenovirus (Adv): 400 μL/mL medium. Cells were harvested 3 days postinfection. ( E ) Protein levels of SPP1, SCD1, and CHAC1 in primary hepatocytes under 100 μM free FA treatment. Primary hepatocytes were infected with Adv overexpressing GFP or Chac1 . Adv: 400 μL/mL medium. Cells were harvested 3 days postinfection. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Journal: The Journal of Clinical Investigation

Article Title: Hepatic glutathione depletion ameliorates MASLD through selective protein oxidation and inhibition of lipogenesis

doi: 10.1172/JCI197556

Figure Lengend Snippet: ( A ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice under chow diet. Expression was assessed by qRT-PCR ( n = 9–11). Mice were fed a chow diet and were sacrificed 8 weeks after infection with AAV. ( B ) Relative expression levels of key genes controlling lipogenesis, β-oxidation, gluconeogenesis, and fibrosis in livers of mice after 10 weeks on HFD. Expression was assessed by qRT-PCR ( n = 7). Mice were fed an HFD for 2 weeks before infection with AAV and were sacrificed 8 weeks later. ( C ) Protein levels of SREBP-1c, SCD1, and CHAC1 in livers of mice under HFD. ( D ) Relative expression levels of Chac1 -regulated genes in primary hepatocytes under 100 μM free fatty acid (FA) treatment. Expression was assessed by qRT-PCR ( n = 3). Adenovirus (Adv): 400 μL/mL medium. Cells were harvested 3 days postinfection. ( E ) Protein levels of SPP1, SCD1, and CHAC1 in primary hepatocytes under 100 μM free FA treatment. Primary hepatocytes were infected with Adv overexpressing GFP or Chac1 . Adv: 400 μL/mL medium. Cells were harvested 3 days postinfection. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Immunoblotting was achieved using the indicated antibodies: phospho-IR Y1152/Y1153 (3024; Cell Signaling Technology [CST]), IR (sc-711), phospho-AKT S473 (4060; CST), phospho-AKT T308 (4056; CST), AKT (4685; CST), phospho-IR T1150 (NBP2-60777; Novus Biologicals), phospho-IRS1 S1097 (2385; CST), phospho-JNK (4668; CST), JNK (9252; CST), CHAC1 (ab76386, Abcam), Vinculin (MAB3574, Sigma), FASN (ab22759, Abcam), SCD1 (2438, CST), total OXPHOS (ab110413, Abcam), NOTCH1 (3608, CST), NOTCH2 (5732, CST), HES1 (sc-166410, Santa Cruz Biotechnology), SPP1 (AF808, R&D Systems), and β-tubulin (2146; CST).

Techniques: Expressing, Quantitative RT-PCR, Infection