aav serotype 2 cmv7 dio sacas9  (Vector Biolabs)

Journal: Molecular Metabolism

Article Title: Deubiquitinating enzyme USP2 regulates brown adipose tissue thermogenesis via controlling EBF2 stabilization

doi: 10.1016/j.molmet.2025.102139

Figure Lengend Snippet: Overexpression of Usp2 in BAT ameliorates HFD-induced obesity and metabolic dysfunction. A. Schematic diagram of the in-situ injection of AAV-OeUsp2 into BAT for the establishment of the Usp2 BOE model, followed by HFD feeding. B. Body weight of Usp2 BOG and control mice housed at RT (22 °C). Usp2 BOE vs control, ( n = 5 vs 6). C. Lean mass and fat mass. Usp2 BOE vs control, ( n = 7 vs 9). D. The weight of indicated tissues ( n = 5). E. Rectal temperature of Usp2 BOE and control mice during acute cold exposure ( n = 8). F. Representative infrared images of Usp2 BOE and control mice following 4 h of acute cold exposure. G. Representative H&E staining of adipose tissue and liver in Usp2 BOE and control mice (Scale bar, 100 μm). H. Quantitative PCR analysis of thermogenic genes in BAT of Usp2 BOE and control mice. Usp2 BOE vs control, ( n = 4 vs 5). I. Representative Ucp1 immunostaining of BAT in Usp2 BOE and control mice (Scale bar, 100 μm). J-L. Oxygen consumption rate ( J ), carbon dioxide production rate ( K ) and heat production rate ( L ) of Usp2 BOE and control mice over 24-hour period monitored at RT (22 °C) ( n = 4). M-O. Average oxygen consumption rate ( M ), average carbon dioxide production rate ( N ) and average heat production rate ( O ) at day time or night time during the 24-hour of monitoring as in (J–L). P. Insulin tolerance test (ITT) results of mice after 8 weeks of HFD feeding ( n = 7). Q. Analysis of ITT results in P. R. Glucose tolerance test (GTT) results of mice after 8 weeks of HFD feeding ( n = 6). S. Analysis of GTT results in R. For statistical analysis, two-way ANOVA was performed in B, E, J-L, P and R. Unpaired, two-tailed t -tests were performed in C, D, H, M−O, Q and S. (∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001). Each point represents a biological replicate. Data were presented as the mean ± S.E.M.

Article Snippet: Adipocyte-specific adeno-associated virus 2/9 (AAV Serotype 2/9, adiponectin promoter)-mediated overexpression of FLAG-tagged mouse Usp2, control (GFP), shRNA targeting Usp2, and scrambled control (NC) were constructed, amplified, and purified by OBiO Technology (Shanghai, China).

Techniques: Over Expression, In Situ, Injection, Control, Staining, Real-time Polymerase Chain Reaction, Immunostaining, Two Tailed Test

Journal: Molecular Metabolism

Article Title: Deubiquitinating enzyme USP2 regulates brown adipose tissue thermogenesis via controlling EBF2 stabilization

doi: 10.1016/j.molmet.2025.102139

Figure Lengend Snippet: EBF2 overexpression attenuates USP2 deficiency-potentiated thermogenic dysfunction. A. Schematic illustration of the establishment of three groups: AAV-NC + Adv-GFP vs AAV-shUsp2 + Adv-GFP vs AAV-shUsp2 + Adv-EBF2, n = 6 vs 5 vs 5. Image created with BioRender.com . B. Tissue weight of BAT. C. Tissue weight of iWAT and eWAT. D. Representative H&E staining of BAT. E. Representative UCP1 immunostaining of BAT. F. Immunoblot analysis of UCP1. G. Quantitative PCR analysis of thermogenic genes of BAT ( n = 5 vs 5 vs 5). H-J. Oxygen consumption rate ( H ), carbon dioxide production rate ( I ) and heat production rate ( J ) of mice over 24-hour period monitored at RT (22 °C) ( n = 4 vs 4 vs 4). K-M. Average oxygen consumption rate ( K ), average carbon dioxide production rate ( L ) and average heat production rate ( M ) at day time or night time during the 24-hour of monitoring as in (H–J). For statistical analysis, two-way ANOVA was performed in H-J. Unpaired, two-tailed t -tests were performed in B, C, G and K-M. (∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001). Each point represents a biological replicate. Data were presented as the mean ± S.E.M.

Article Snippet: Adipocyte-specific adeno-associated virus 2/9 (AAV Serotype 2/9, adiponectin promoter)-mediated overexpression of FLAG-tagged mouse Usp2, control (GFP), shRNA targeting Usp2, and scrambled control (NC) were constructed, amplified, and purified by OBiO Technology (Shanghai, China).

Techniques: Over Expression, Staining, Immunostaining, Western Blot, Real-time Polymerase Chain Reaction, Two Tailed Test