Journal: International Journal of Molecular Sciences
Article Title: Orphan Nuclear Receptor ERRγ Is a Transcriptional Regulator of CB1 Receptor-Mediated TFR2 Gene Expression in Hepatocytes
Figure Lengend Snippet: ERRγ increases TFR2 mRNA and protein levels. ( A – C ) Q-PCR analysis showing ERRγ, TFR1, and TFR2 mRNA levels. HepG2 cells were transfected with vectors expressing ERRα ( A ), ERRβ ( B ), and ERRγ ( C ) for 48 h. ( D ) Q-PCR analysis showing ERRγ, TFR1, and TFR2 mRNA levels. AML12 cells were transfected with vector expressing ERRγ for 48 h. ( E ) Q-PCR analysis showing TFR2 mRNA levels. HepG2 cells and AML12 cells were treated with GSK4716 (10 μM) for 12 h. ( F ) Q-PCR analysis showing ERRγ and TFR2 mRNA levels in the liver. Ad-GFP and Ad-ERRγ were injected via tail veins into male C57BL/6J mice ( n = 3 per group). Mice were then sacrificed at day 6. ( G , H ) Western blot analysis ( G ) and graphical representation ( H ) showing TFR2 protein levels. Huh7 cells were transfected with vector expressing ERRγ (+: 1 μg, ++: 3 μg) for 48 h. ( I , J ) Western blot analysis ( I ) and graphical representation ( J ) showing TFR2 protein levels. AML12 cells were transfected with a vector expressing ERRγ (+: 3 μg) for 48 h. Experiments were performed in duplicate ( G – J ) or triplicate ( A – F ) and repeated at least twice. Error bars show ± SD. * p
Article Snippet: Cell Culture, Transfection and Luciferase Assay HepG2 (human liver cancer cell line; ATCC, Manassas, VA, USA) cells and Huh7 (human hepatoma cell line, ATCC) cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, high glucose, Welgene, Gyeongsangbuk-do, Korea) and RPMI 1640 medium (Welgene) supplemented with 10% fetal bovine serum and 1% antibiotics (penicillin-streptomycin).
Techniques: Polymerase Chain Reaction, Transfection, Expressing, Plasmid Preparation, Injection, Mouse Assay, Western Blot