Journal: PLoS ONE

Article Title: Regulation of gene transcription by thyroid hormone receptor β agonists in clinical development for the treatment of non-alcoholic steatohepatitis (NASH)

doi: 10.1371/journal.pone.0240338

Figure Lengend Snippet: (A) Illustration of the in vitro , hepatic cell-based differential gene expression assay design. (B) THRB and THRA RNA levels were quantified by RT-qPCR in HepG2 (n = 3), Huh-7, (n = 3), and PHH (n = 5) cells. Mean RQ values ± SEM are reported with means annotated within the bars. (C) Huh-7 cells were treated with increasing doses of T3 (n = 2), GC-1 (n = 2), or MGL-3196 (n = 2) for 24 hrs. ANGPLT4 , CPT1A , and DIO1 RNA levels were quantified by RT-qPCR and dose-response curves were generated for each gene-compound combination. Mean EC 50 values (red bar) and individual replicate EC 50 values (black symbols) are reported. (D) Huh-7 cells were treated with increasing doses of T3 (black), GC-1 (red), MGL-3196 (green), VK2809A (solid blue), or VK2809 (dashed blue) for 24 hrs. CPT1A RNA levels were quantified by RT-qPCR. Representative mean RQ values at each compound concentration and fitted dose-response curves are reported. (E) PHH were treated with increasing doses of T3 (black), GC-1 (red), MGL-3196 (green), VK2809A (solid blue), or VK2809 (dashed blue) for 24 hrs. THRSP RNA levels were quantified by RT-qPCR. Representative mean RQ values at each compound concentration and fitted dose-response curves are reported. (F) EC 50 values for every test compound were calculated from dose-response curves generated from the TR-FRET THRβ, luciferase (Luc) reporter THRβ, Huh-7 differential gene expression (RQ), and PHH RQ assays (data reported in Tables and ). Mean EC 50 values ± SEM are reported.

Article Snippet: Transporter certified human hepatocytes (PHH) were obtained from BioIVT (Lot: JEL, F00995-TCERT).

Techniques: In Vitro, Gene Expression, Quantitative RT-PCR, Generated, Concentration Assay, Luciferase

Journal: Redox Biology

Article Title: Vitamin E treatment in NAFLD patients demonstrates that oxidative stress drives steatosis through upregulation of de-novo lipogenesis

doi: 10.1016/j.redox.2020.101710

Figure Lengend Snippet: αT inhibits de novo lipogenesis (DNL) in vitro . HepG2 cells were incubated with normal growth medium (NGM, white) or high glucose medium (HGM, 25 mmol/L glucose, blue shades) for 48 h. Incorporation of 14 C-Glucose into lipids was used as a measure of DNL. Concurrent incubation of cells with 25–100 μmol/L αT decreased cellular triglycerides (A) and DNL (C). αT (100 μmol/L) inhibits HGM-induced upregulation of fatty acid synthase (FASN) and stearoyl-CoA desaturase (SCD) mRNA expression in HepG2 (D) and primary human hepatocytes (PHH, n = 3 donors) (E). PHH experiments were normalized within each donor. All n = 3 experiments; A-C mean ± SD, D-E mean ± SEM. All samples were compared to HGM solvent control (S·C.). Bars not sharing the same letter are significantly different p < 0.05. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Primary human hepatocytes (Lonza, USA, PHH) were cultivated in Lonza HBM TM Basal Media with HCM SingleQuots (Lonza, USA).

Techniques: In Vitro, Incubation, Expressing, Solvent, Control