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: 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

Journal: Cell Reports Medicine

Article Title: Hepatitis Delta Virus Acts as an Immunogenic Adjuvant in Hepatitis B Virus-Infected Hepatocytes

doi: 10.1016/j.xcrm.2020.100060

Figure Lengend Snippet:

Article Snippet: Primary human hepatocytes (Cryopreserved) , Lonza , HUM4012.

Techniques: Staining, Recombinant, Amplification, Virus, Enzyme-linked Immunosorbent Assay, RNAscope, Multiplex Assay, Clone Assay, In Situ, Plasmid Preparation, Software

Journal: EMBO Molecular Medicine

Article Title: A critical role for HNF4α in polymicrobial sepsis-associated metabolic reprogramming and death

doi: 10.1038/s44321-024-00130-1

Figure Lengend Snippet: ( A ) Experimental setup porcine sepsis model. By intraperitoneal installation of 3 g/kg of autologous feces, the animals were permitted to develop sepsis until reaching a state of severe hypotension characterized by a mean arterial pressure (MAP) of ≤50 mmHg. Once severe hypotension, defined as a MAP ranging between 45 and 50 mmHg, was achieved, it was maintained for a duration of one hour. Around 18 h after sepsis initiation, liver samples were collected for RNA-Seq analysis ( n = 3 sham, n = 9 sepsis; biological replicates). ( B ) Enrichr analysis of the genes downregulated (Padj < 0.05, LFC < 0) in pig sepsis. Differential genes were identified by DESeq2 (using Wald test). The upstream regulators are ordered by their enrichment P -value (derived from Fisher’s exact test (Hypergeometric test)). ( C ) HOMER transcription factor motif enrichment 1000 bp upstream of TSS from the genes downregulated in pig sepsis. Differential genes were identified by DESeq2 (using Wald test). P -values were derived from Fisher’s exact test (Hypergeometric test)). ( D ) At 2 weeks, Alb-uPA +/+ -SCID mice were injected with human hepatocytes into their spleens, enabling migration to the liver and repopulation of empty niches. After several weeks, mice underwent sham or CLP surgery, and livers were isolated 24 h later. ( E ) Heatmaps displaying the expression levels of HNF4α-dependent genes (genes differentially expressed in Hnf4a Liver-i-KO sham relative to Hnf4a flfl sham, with Padj < 0.05) in mouse liver 24 h post-CLP, pig sepsis liver and humanized mice liver 24 h post-CLP. We selected HNF4α target genes known to be downregulated (Padj < 0.05) in mouse liver 24 h post-CLP, focussing on the ones mentioned in the paper. Differential genes were identified by DESeq2 (using Wald test). Expression levels were quantified by RT-qPCR relative to Hprt and Rpl , and log transformed. For the humanized mice, human-specific and mouse-specific primers were used to investigate the response in human and mouse hepatocytes, respectively.

Article Snippet: Briefly, the mice were generated by transplanting homozygous Alb-uPA +/+ -SCID mice (which suffer from spontaneous and chronic death of hepatocytes) with 0.7 × 10 6 cryopreserved primary human hepatocytes (donor C342, Lonza), via intrasplenic injection.

Techniques: RNA Sequencing, Derivative Assay, Injection, Migration, Isolation, Expressing, Quantitative RT-PCR, Transformation Assay