Journal: Molecular Therapy

Article Title: CasRx-based Wnt activation promotes alveolar regeneration while ameliorating pulmonary fibrosis in a mouse model of lung injury

doi: 10.1016/j.ymthe.2024.09.008

Figure Lengend Snippet: AAV6 shows high infection efficiency and specificity in lung epithelial cells (A) Schematic illustration of IT injection of AAV-GFPs followed by lung harvest at day14 for fluorescence-activated cell sorting (FACS) and immunofluorescence (IF) staining analysis. (B) The proportion of GFP + cells in live cells in AAV5-GFP-, AAV6-GFP-, and AAV9-GFP-infected lungs by flow cytometry analysis. n = 4 mice per group. (C) The mean GFP intensity in AAV5-GFP-, AAV6-GFP-, and AAV9-GFP-infected lungs by flow cytometry analysis, normalized by that in AAV9 group. n = 4 mice per group. (D) Representative images of GFP immunofluorescence in AAV5-GFP-, AAV6-GFP-, and AAV9-GFP-infected lung sections. Scale bar, 200 μm. (E) The proportion of GFP + cells in different cell types by flow cytometry analysis. n = 4 mice per group. (F and G) Representative images of IF staining of SPC (AT2 cells), RAGE (AT1 cells), CC10 (club cells), and acetylated-tubulin (ciliated cells) in AAV6-GFP infected lung sections. Scale bar, 40 μm. (B, C, and E) Mean ± SEM and p values were analyzed by one-way ANOVA. Data are representative of three independent experiments.

Article Snippet: AAV5-GFP, AAV6-GFP and AAV9-GFP were purchased from PackGene Biotech.

Techniques: Infection, Injection, Fluorescence, FACS, Immunofluorescence, Staining, Flow Cytometry

Journal: Theranostics

Article Title: Cardiomyocyte-derived OTUD7B promotes cardiac hypertrophy by deubiquitinating SERCA2a

doi: 10.7150/thno.129105

Figure Lengend Snippet: Cardiomyocyte-specific OTUD7B overexpression promotes cardiac hypertrophy by deubiquitinating SERCA2a at K628. (A) Schematic diagram of the TAC-induced mouse model. WT mice were injected with AAV9 cardiomyocyte-specific overexpressing SERCA2a WT or SERCA2a K628R , and OTUD7B OE or empty vector (EV) by the tail vein. After 2 weeks, mice were subjected to TAC. After 4 weeks, the cardiac function of mice was assessed using echocardiography. Mice were then euthanized, and samples were harvested. (B) Representative left ventricular M-mode echocardiographic images. (C-D) Values of ejection fraction (C) and fractional shortening (D). EF, ejection fraction; FS, fractional shortening; n = 6. (E) Representative whole heart images. (F) The ratio of heart weight to body weight (HW/BW). n = 6. (G) The ratio of heart weight to tibial length (HW/TL). n = 6. (H) Representative HE-stained images of cardiac tissue sections. (I-J) Representative images (I) and quantification (J) of wheat germ agglutinin (WGA)-stained cardiac tissue sections (n = 6). (K-L) Representative images (K) and quantification (L) from Masson's trichrome-stained cardiac tissue sections (n = 6). (M) Serum atrial natriuretic peptide (ANP) levels were detected using ELISA kits (n = 6). (N-P) mRNA levels of Myh7 (N), Nppa (O), and Nppb (P) in heart tissues (n = 6). Signal intensities were adjusted using Actb as the normalization control.

Article Snippet: Animals were maintained for 4 weeks after TAC or sham surgery. (3) OTUD7B (OTUD7B OE ) and SERCA2a (SERCA2a WT or SERCA2a K628R ) cardiomyocyte-specific overexpression was achieved using recombinant adeno-associated virus serotype 9 (AAV9) vectors driven by the cTNT promoter (Genechem, Shanghai, China).

Techniques: Over Expression, Injection, Plasmid Preparation, Staining, Enzyme-linked Immunosorbent Assay, Control