aav9 paav cmv hi cre gfp wpre addgene addgene  (Addgene inc)

Journal: The Journal of Clinical Investigation

Article Title: Ventromedial hypothalamic primary cilia control energy and skeletal homeostasis

doi: 10.1172/JCI138107

Figure Lengend Snippet: (A) Primary cilia were visualized by ACIII (green) in the VMH of WT and IFT88-KOSF-1 mice. Scale bar: 10 μm. (B) Length and percentage of the VMH cilia shown in A. **P < 0.01; ***P < 0.001, Student’s t test. (C) Left: weekly body weight of WT and IFT88-KOSF-1 mice fed on NC. Right: representative photographs of 18-week-old male mice on NC diet. *P < 0.05, 2-way ANOVA. (D) Left: fat and lean mass were measured by NMR. Right: representative DEXA image. **P < 0.01, Student’s t test. (E) Representative s.c.WAT and gWAT from WT and IFT88-KOSF-1 mice. Scale bars: 0.5 mm. (F and G) H&E staining (F) and adipocyte cell surface (G) of gWAT. Adipocyte cell surface measured by ImageJ. Scale bar: 50 μm. ***P < 0.001, 2-way ANOVA. (H) Serum leptin levels in WT and IFT88-KOSF-1 mice at 14 week of age. **P < 0.01, Student’s t test. (I) Left: schematic showing bilateral viral injection into VMH. Right: schematic for AAV2-GFP and AAV2-Cre-GFP constructs used for viral injection (upper) and validation images of primary cilia KD in VMH (lower). Scale bars: 0.1 mm (yellow); 10μm (white). (J) Weekly body weight after injection with AAV2-GFP or AAV2-Cre-GFP into the VMH. ***P < 0.001, 2-way ANOVA. (K) Left: body compositions 10 weeks after injection of AAV2-GFP or AAV2-Cre-GFP. Right: representative DEXA images. ***P < 0.001, Student’s t test. Number of animals examined is expressed in parentheses in each graph. Results are expressed as mean ± SD.

Article Snippet: AAV expressing Cre-GFP (catalog 105545, AAV2-Cre-GFP) and GFP (catalog 105530, AAV2-GFP) were purchased from Addgene.

Techniques: Staining, Injection, Construct, Biomarker Discovery

Journal: The Journal of Clinical Investigation

Article Title: Ventromedial hypothalamic primary cilia control energy and skeletal homeostasis

doi: 10.1172/JCI138107

Figure Lengend Snippet: (A–D) DEXA analyses were performed at indicated ages and measured bone parameters. BMD (A), BMC (B), BV (C), and BA (D) levels of WT and IFT88-KOSF-1 mice. *P < 0.05; **P < 0.01, 2-way ANOVA. (E–H) Correlation between serum NE and BMD (E), BMC (F), BV (G), and BA (H) respectively. For correlation testing, linear regression models in R were utilized and R2 values were reported. (I) Femur NE level of WT and IFT88-KOSF-1 littermates at 16 weeks of age. ***P < 0.001, Student’s t test. (J) Representative μCT images of the femurs. The 12-week old WT and IFT88-KOSF-1 littermates were used for analyses. (K) BMD, BV/TV, BS, Tb.Th., Tb.N., and Tb.Sp. were analyzed using μCT from J. *P < 0.05; **P < 0.01, Student’s t test. (L) Representative μCT images of femurs in control (AAV2-GFP) and viral-mediated VMH-specific primary cilia KD (AAV2-Cre-GFP) mice. (M) BMD, BV/TV, BS, Tb.Th., Tb.N., and Tb.Sp. were analyzed using μCT from L. *P < 0.05, Student’s t test. Number of animals examined is expressed parentheses in each graph. Data are expressed as mean ± SD.

Article Snippet: AAV expressing Cre-GFP (catalog 105545, AAV2-Cre-GFP) and GFP (catalog 105530, AAV2-GFP) were purchased from Addgene.

Techniques: Control