F-box only protein 2 exacerbates non-alcoholic fatty liver disease by targeting the hydroxyl CoA dehydrogenase alpha subunit

Figure 6 F-box only protein 2 ubiquitinated and degraded the hydroxyl CoA dehydrogenase alpha subunit. A: The hydroxyl CoA dehydrogenase alpha subunit (HADHA) ubiquitination in HEK293T cells; B-D: Western blot of F-box only protein 2 (FBXO2) and HADHA expression in the His-Vector group vs His-FBXO2 group (B), Sh-NC group vs Sh-FBXO2 group (C), and adeno-associated virus (AAV)-His-vector mice vs AAV-His-FBXO2 mice (D); E: Western blot of FBXO2 and HADHA expression in HepG2 cells. HepG2 cells were transfected with His-FBXO2 plasmids for 24 h, followed by incubation with or without MG132 for 24 h; F: Western blot of FBXO2 and HADHA expression in HepG2 cells. HepG2 cells were transfected with FBXO2 adenovirus for 48 h, followed by incubation with cycloheximide (CHX) for 0, 4, or 8 h before sample collection; G-J: HepG2 cells were transfected with His-FBXO2 plasmids and Myc-HADHA plasmids to overexpress FBXO2 and HADHA, respectively, followed by treatment with oleate for 24 h; representative images of Oil Red O staining (G), triglyceride (H) and total cholesterol (I) levels; expression analysis of genes associated with lipogenesis, fatty acid uptake, and oxidation as detected using quantitative reverse transcription polymerase chain reaction (J). FBXO2: F-box only protein 2; HADHA: Hydroxyl CoA dehydrogenase alpha subunit; ^aP < 0.05, ^bP < 0.01, ^cP < 0.001. AAV: Adeno-associated virus; OA: Oleate; TG: Triglyceride; T-Cho: Total cholesterol; FASN: Fatty acid synthase; PPAR: Peroxisome proliferator-activated receptor; CD36: Cluster of differentiation 36; ACOX1: Acyl-CoA oxidase 1; ECH1: Enoyl coenzyme A hydratase 1; MCAD: Medium-chain acyl-coenzyme A dehydrogenase; NC: Normal control.