Journal: PLoS ONE

Article Title: Strategies to Rescue the Consequences of Inducible Arginase-1 Deficiency in Mice

doi: 10.1371/journal.pone.0125967

Figure Lengend Snippet: (A) Maps (not to scale) of two AAV constructs used and the packaging serotypes (AAV8 and rh10). ITR, inverted terminal repeat; TBG, thyroxine-binding globulin; CB7-CI, hybrid cytomegalovirus enhancer/chicken β-actin promoter (CB7), along with a chicken β-actin intron (CI); WPRE, woodchuck hepatitis virus post-transcriptional regulatory element; bGH-pA, bovine growth hormone polyA adenylation signal sequence; rBG, rabbit beta-globin polyA adenylation signal sequence. (B) Western blot indicating level of expression in livers of 4 separate mice (lanes 1–4) of AAV-expressed Arg1-eGFP relative to WT Arg1 and tubulin loading control post-tamoxifen (Day +13;4-week old mice injected with 2x10 11 gc of the AAV8 construct shown in panel A) on Day -4. Lane 5 represents normal level of expression of Arg1 (and tubulin) in a mouse not injected with tamoxifen. In order to observe sufficient signal of the inducible Arg1 knockout band of lanes 1–4, the signal becomes over-saturated when detecting normal endogenous levels of Arg1 (lane 5). (C) Images of liver sections (i-iii) and dissociated hepatocytes (iv, v) from AAV8.TBG.PI.Arg1-eGFP.WPRE.bGH injected 4-week old mice 1 week after virus administration. Strong fluorescence signal from the fusion transgene is observed in the cytoplasm of some periportal hepatocytes (ii,iii) and also in dissociated cells (v) with corresponding phase contrast images (i, iv).

Article Snippet: The first was driven by the liver-selective thyroxine-binding globulin (TBG) promoter with a chimeric intron from Promega (PI), encoding the 5′-donor site from the first intron of the human β-globin and the branch and 3′-acceptor site from the intron located between the leader and body of an immunoglobulin gene heavy chain variable region.

Techniques: Construct, Binding Assay, Sequencing, Western Blot, Expressing, Injection, Knock-Out, Fluorescence

Journal: Molecular Metabolism

Article Title: Membrane-associated ring–CH–type finger 2 protects against metabolic dysfunction-associated fatty liver disease by targeting fatty acid synthase

doi: 10.1016/j.molmet.2025.102137

Figure Lengend Snippet: Overexpression of hepatic MARCH2 ameliorates MAFLD in ob/ob mice. (A) The rAAV8 with TBG promoter-driven MARCH2 (AAV-MARCH2) or GFP (AAV-GFP) were delivered by tail vein injection to 8-week-old ob/ob mice and fed with a chow diet for another 7 weeks. This schema diagram was created by Figdraw (authorization code: TTPIO86bdd). Body weight (B), morphology of the mice and liver (C), GTT (D), ITT (E), area under curve(AUC) of GTT(F), AUC of ITT(G), liver weight (H), ratios of liver weight to body weight (I), liver HE staining (J), liver Oil Red O staining (K), liver TG contents (L), liver MARCH2 mRNA expression (M), and FASN protein levels (N) of AAV-MARCH2 and AAV-GFP mice were detected. n = 6–8 mice per group, and the data were expressed as Mean ± SEM. Two-tailed unpaired student’s t -test were applied.

Article Snippet: The rAAV8 with thyroxine binding globulin (TBG) promoter-driven MARCH2 coding sequence (AAV-MARCH2) or GFP control (AAV-GFP) were generated by GenePharma (Shanghai, China).

Techniques: Over Expression, Injection, Staining, Expressing, Two Tailed Test