Journal: bioRxiv

Article Title: Angiotensin II Type 2 Receptor Potentiates Skeletal Muscle Satellite Cell Differentiation via the GSK3β/β-catenin Pathway

doi: 10.1101/2022.09.30.510328

Figure Lengend Snippet: (A) SC-CAG-AT2R animal model. Left panel: In SC-CAG-AT2R (Pax7 CreER/+ ; CAG-CAT flox -AT2R; R26 LSL-EYFP ) mice, neither AT2R nor EYFP is expressed due to the presence of stop codons upstream of AT2R or EYFP. CreER is expressed specifically in SCs under the control of Pax7 promoter but inactive in the absence of tamoxifen (Tmx). Right panel: Upon tamoxifen administration, CreER becomes active and stop codons are removed via Cre-mediated recombination, allowing AT2R or EYFP to be expressed specifically in SCs. (B) Various tissues and SCs were harvested from control (Pax7CreER/+; R26 LSL-EYFP ) and SC-CAG-AT2R mice ±Tmx administration. AT2R mRNA expression was quantified by qRT-PCR. N=4; mean±SEM; *p<0.05. (C) Primary SCs were collected from SC-CAG-AT2R and control SC-EYFP mice, cultured in vitro , and induced to differentiate for 2 days. Cells were harvested before (d0) and after 2 days of differentiation, and gene expression was analyzed by qRT-PCR. N=5; mean±SEM; *p<0.05; **p<0.01. (D, E) SCs were collected as in (C), and myofiber formation was visualized in bright field (top panels in D) and by EYFP fluorescence (bottom panels in D), and myofiber diameter was quantified (E). N=4; mean±SEM; *p<0.05. (F) SCs were collected as in (A), and AT2R and myogenin protein expression was measured by immunoblotting.

Article Snippet: The amplified sequence was subcloned into CAG-lox-CAT-lox vector(gift from Jeffrey Robbins; Addgene plasmid # 53959) using BamHI cloning site.

Techniques: Animal Model, Expressing, Quantitative RT-PCR, Cell Culture, In Vitro, Fluorescence, Western Blot

Journal: bioRxiv

Article Title: Angiotensin II Type 2 Receptor Potentiates Skeletal Muscle Satellite Cell Differentiation via the GSK3β/β-catenin Pathway

doi: 10.1101/2022.09.30.510328

Figure Lengend Snippet: (A) Map of mouse AT2R gene (top) and transgenic vector for CAG-CAT flox -AT2R mice (bottom). (B) Southern blotting of AT2R gene in four CAG-CAT flox -AT2R founder lines (lines 888, 898, 900 and 902) with the probe indicated in (A). Genomic DNA of these animals were digested with BamHI. The location of the restriction enzyme digestion sites and the predicted DNA fragment sizes are shown (A). Copy numbers from each line are shown at the bottom. (C) Primary SCs were collected from SC-CAG-AT2R, SC-EYFP and negative control Pax7 +/+ ; R26 LSL-EYFP/+ mice, and bright field and EYFP fluorescence images are shown.

Article Snippet: The amplified sequence was subcloned into CAG-lox-CAT-lox vector(gift from Jeffrey Robbins; Addgene plasmid # 53959) using BamHI cloning site.

Techniques: Transgenic Assay, Plasmid Preparation, Southern Blot, Negative Control, Fluorescence