Journal: Molecular Therapy. Nucleic Acids

Article Title: MicroRNA199a-Based Post-transcriptional Detargeting of Gene Vectors for Hepatocellular Carcinoma

doi: 10.1016/j.omtn.2018.08.016

Figure Lengend Snippet: Expression Pattern of miRNA199a The expression levels of miRNA199a in primary hepatocytes HepaRG and a panel of HCC (A) and non-HCC cell lines (B) as assayed by real-time qPCR. The copies of miRNA199a were normalized against those of the RNU6 control and represented as the number of copies per 1,000 control (n > 3 for all cell lines except HUM4150).

Article Snippet: Cryopreserved primary human hepatocytes (HUM4150) and NoSpin HepaRG (NSHPRG) cells were obtained from Lonza (Sydney, Australia) and maintained as per the manufacturer’s protocol.

Techniques: Expressing, Control

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