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. 2023 Dec 8;15(24):5046. doi: 10.3390/nu15245046

Figure 5.

Figure 5

Combined treatment of adiponectin and metformin ameliorates the regulation of FA synthesis in low-B12 hepatocytes via increased AMPK activation: (A) The mRNA expression of AdipoR1 (i) and R2 (ii) and master regulator of lipogenesis SREBF1 (C) (i) as well as enzymes regulating fatty acid synthesis, including ACC (ii), FASN (iii) and ELOVL6 (iv), TG synthesis (D), including SCD (i), DGAT1 (ii) and DGAT2 (iii) and cholesterol LDLR (E), were normalized to 18S rRNA endogenous control (Applied Biosystems, Knutsford, UK). (B) Protein levels of pAMPKα (i) and pACC (ii) following combined or sequential treatment of adiponectin and metformin and normalized to total AMPK and ACC, respectively. (F) Total intracellular levels of triglycerides quantified in hepatocytes using the TG kit (ab65336) (Abcam Plc, Cambridge, UK) and normalized per milligram protein under each B12 condition. (G) Levels of synthesized TGs in hepatocytes assessed with the radioactive flux assay and quantified as disintegrations per minute (DPM) using the scintillation counter and normalized per milligram (mg) protein. The data is representative of mean ± SEM (n = 6), and * represents significance compared to controls B12 (500 nM) and low B12 (25 pM), $—compared to controls B12 (500 nM) in metformin only or both metformin and adiponectin and &—compared to low B12 (25 pM) in both metformin and adiponectin, respectively; * p < 0.05, ** p < 0.01, *** p < 0.001; $ < 0.05, $$ < 0.01, $$$ < 0.001; && < 0.01, &&& < 0.001.