Fig 1: Genistein improves the oxidative capacity of skeletal muscle in subjects with obesity and insulin resistance (HOMA index greater than 2.5) after 2 months of treatment. (A, B) Metabolomic analysis of the compounds derived from the benzoate metabolism of bacteria from the gut microbiota assessed by mass spectrometry at basal (B) and final (F) assessment. (C) Hierarchical clustering heat map showing the relative intensity of the 376 differentially expressed genes (206 upregulated and 170 downregulated) in the GTG versus the PG. Analysis using Partek genomics suite; blue indicates downregulated expression and red indicates upregulated expression, with intensity of color reflecting intensity of upregulation or downregulation. (D) Comparison of the percentage of upregulated and downregulated genes associated with fatty acid oxidation in the GTG versus the PG. (E) Microscopic analysis of SDH staining (up) and BODIPY 493/503 staining (down) showing mitochondrial activity and lipid accumulation, respectively, in sections of OCT-embedded skeletal muscle biopsies from the GTG and PG. The nuclei were stained with DAPI. The bar measures 200 µm in size. (F) Densitometric quantitation of SDH staining and (G) BODIPY staining using ImageJ. (H) Immunoblotting analysis of total ACC, p-ACC, total AMPK and p-AMPK of skeletal muscle biopsies of obese patients in GTG and PG. (I) Densitometric quantitation of p-ACC/total ACC and (J) p-AMPK/total AMPK. values are given as mean±SEM (n=10). Differences between the two groups were evaluated by unpaired t-test. *P<0.05, ***P<0.001. ACC, acetyl-CoA carboxylase; AMPK, 5'-adenosine monophosphate-activated protein kinase; BODIPY, boron-dipyrromethene; DAPI, 4',6-diamidino-2-phenylindole; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GTG, genistein-treated group; HOMA, Homeostatic Model Assessment; OCT, Optimal Cutting Temperature reagent; p-ACC, Phospho-Acetyl-CoA Carboxylase; p-AMPK, Phospho-AMP-activated protein kinase; PG, placebo group; PPi, Pyrophosphate; SDH, succinate dehydrogenase.