recombinant aav vectors  (Addgene inc)

Journal: Inflammation

Article Title: IRF8 and FOXM1 Regulation in COPD Progression: Impacts on Inflammation and Senescence

doi: 10.1007/s10753-026-02474-x

Figure Lengend Snippet: Knockdown of IRF8 alleviates lung injury in COPD mice. COPD model mice were intratracheally administered with corresponding adeno-associated virus to interfere with IRF8 expression. ( A ) Lung index calculated as lung weight/body weight. ( B - E ) Assessment of emphysema in lung tissue by H&E staining. B: Representative H&E-stained images; C: Mean alveolar number; D: Mean linear intercept; E: Airway inflammation score based on inflammatory cell infiltration. ( F ) Real-time PCR analysis of IRF8 expression in lung tissue. ( G ) Western blot analysis of IRF8 protein levels in lung tissue. ( H - L ) Giemsa staining and quantification of total leukocytes ( H ), neutrophils ( I ), Eosinophil ( J ), macrophages ( K ), and lymphocytes ( L ) in bronchoalveolar lavage fluid (BALF). ( M - O ) ELISA detection of TNF-α (M), IL-1β ( N ), and IL-1α ( O ) levels in lung tissue. N = 6 replicates

Article Snippet: Recombinant adeno-associated virus (AAV) vectors were constructed and produced using the shuttle plasmid pAV-U6-shRNA-CMV-RFP (Vigene Biosciences, pAV100004-KD).

Techniques: Knockdown, Virus, Expressing, Staining, Real-time Polymerase Chain Reaction, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: S-9-PAHSA ameliorates cognitive decline in a type 2 diabetes mouse model by inhibiting oxidative stress and apoptosis via CAIII modulation

doi: 10.3389/fnmol.2025.1617543

Figure Lengend Snippet: The knockdown of CAIII in the hippocampus and the effects of SP during the administration. (A) Overview of the experimental design; (B–G) protein expression and statistical analysis of CAIII in the hippocampus (B,E) , cortex (C,F) and the remaining brain (excluding the hippocampus and cortex) (D,G) of mice following injection with control or CAIII shRNA, n = 5; Body weight (H) , fasting glucose (I) , water intake (J) , and food intake (K) in mice during SP treatment, n = 15; (L,M) IPGTT results and AUC, n = 9; (N,O) IPITT results and AUC, n = 9. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant AAV vectors encoding CAIII shRNA (5′-GGTTCACTGGAATCCAAAGTA-3′) or non-specific control shRNA (5′-CGCTGAGTACTTCGAAATGTC-3′) were synthesized by Shanghai Genechem Co., Ltd. Mice were anesthetized with 1.2% isoflurane and placed on a stereotaxic apparatus (RWD, China) with a heating pad to maintain body temperature at 37°C.

Techniques: Knockdown, Expressing, Injection, Control, shRNA

Journal: Frontiers in Molecular Neuroscience

Article Title: S-9-PAHSA ameliorates cognitive decline in a type 2 diabetes mouse model by inhibiting oxidative stress and apoptosis via CAIII modulation

doi: 10.3389/fnmol.2025.1617543

Figure Lengend Snippet: CAIII-mediated the improvement of SP in the cognitive impairment of T2DM mice. (A) Y-maze test track images from mice, n = 10 ~ 11; The total distance moved (B) , velocity (C) , and alternation (D) for different groups in the Y maze test; (E) Representative tracks of each group on the probe trial day in the Morris water maze test; (F) latency to platform over 5 training days; Escape latency (G) , platform crossings (H) , time at target platform (I) , time in target quadrant (J) , total distance (K) and movement velocity (L) of each group on the probe trial day, n = 8 ~ 10; Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant AAV vectors encoding CAIII shRNA (5′-GGTTCACTGGAATCCAAAGTA-3′) or non-specific control shRNA (5′-CGCTGAGTACTTCGAAATGTC-3′) were synthesized by Shanghai Genechem Co., Ltd. Mice were anesthetized with 1.2% isoflurane and placed on a stereotaxic apparatus (RWD, China) with a heating pad to maintain body temperature at 37°C.

Techniques:

Journal: Frontiers in Molecular Neuroscience

Article Title: S-9-PAHSA ameliorates cognitive decline in a type 2 diabetes mouse model by inhibiting oxidative stress and apoptosis via CAIII modulation

doi: 10.3389/fnmol.2025.1617543

Figure Lengend Snippet: SP ameliorated neuronal injury and apoptosis via CAIII-mediated Bcl-2/Bax and AMPK/Sirt1/PGC1α pathway. Hippocampal ROS (A) , SOD activity (B) , CAT (C), and GSH-Px (D) and H 2 O 2 (E) in mice, n = 4 ~ 5. (F) NeuN immunostaining in hippocampal sections across groups; NeuN-positive neuron percentages in CA1 (G) , CA3 (H) , and DG (I) regions, normalized to the HFD + AAV-Con+vehicle group, n = 4; (J–Q) Western blot analysis of Bcl-2, Bax, Bcl-xl, and cleaved-Caspase 3 (cl-casp3) in mouse hippocampus, n = 3; (R-U) Western blot analysis of p -AMPK, AMPK, Sirt1 and PGC1α, n = 3. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant AAV vectors encoding CAIII shRNA (5′-GGTTCACTGGAATCCAAAGTA-3′) or non-specific control shRNA (5′-CGCTGAGTACTTCGAAATGTC-3′) were synthesized by Shanghai Genechem Co., Ltd. Mice were anesthetized with 1.2% isoflurane and placed on a stereotaxic apparatus (RWD, China) with a heating pad to maintain body temperature at 37°C.

Techniques: Activity Assay, Immunostaining, Western Blot

Journal: Frontiers in Molecular Neuroscience

Article Title: S-9-PAHSA ameliorates cognitive decline in a type 2 diabetes mouse model by inhibiting oxidative stress and apoptosis via CAIII modulation

doi: 10.3389/fnmol.2025.1617543

Figure Lengend Snippet: The effects and mechanism of SP on cell viability, LDH release, and mitochondrial production in PC12 cells. NC, CAIII sh, and CAIII OE cells were exposed to standard medium (Con), high-glucose and high-fat medium (100 mM glucose + 200 μM palmitic acid, G100F200), G100F200 with DMSO, or G100F200 with 60 μM SP for 24 h. (A–D) Cell viability of CAIII sh cells, CAIII OE cells, and NC cells, n = 6; (E–H) LDH release of CAIII sh cells, CAIII OE cells and NC cells, n = 5 ~ 6; (I) Flow cytometric analysis of apoptosis using Annexin V-FITC/PI staining. (J) Quantification of apoptosis rate based on flow cytometry results. SOD activity (K–N) , CAT (O–R) , and GSH-Px (S–V) of CAIII sh cells, CAIII OE cells, and NC cells, n = 6. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant AAV vectors encoding CAIII shRNA (5′-GGTTCACTGGAATCCAAAGTA-3′) or non-specific control shRNA (5′-CGCTGAGTACTTCGAAATGTC-3′) were synthesized by Shanghai Genechem Co., Ltd. Mice were anesthetized with 1.2% isoflurane and placed on a stereotaxic apparatus (RWD, China) with a heating pad to maintain body temperature at 37°C.

Techniques: Staining, Flow Cytometry, Activity Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: S-9-PAHSA ameliorates cognitive decline in a type 2 diabetes mouse model by inhibiting oxidative stress and apoptosis via CAIII modulation

doi: 10.3389/fnmol.2025.1617543

Figure Lengend Snippet: The effects and mechanism of SP on mitochondrial production in PC12 cells. MitoSOX Red immunofluorescence for mitochondrial ROS and quantification in NC (A,B) and CAIII sh (C,D) cells, n = 3. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant AAV vectors encoding CAIII shRNA (5′-GGTTCACTGGAATCCAAAGTA-3′) or non-specific control shRNA (5′-CGCTGAGTACTTCGAAATGTC-3′) were synthesized by Shanghai Genechem Co., Ltd. Mice were anesthetized with 1.2% isoflurane and placed on a stereotaxic apparatus (RWD, China) with a heating pad to maintain body temperature at 37°C.

Techniques: Immunofluorescence

Journal: Frontiers in Molecular Neuroscience

Article Title: S-9-PAHSA ameliorates cognitive decline in a type 2 diabetes mouse model by inhibiting oxidative stress and apoptosis via CAIII modulation

doi: 10.3389/fnmol.2025.1617543

Figure Lengend Snippet: Differentially expressed genes of biological processes and pathways after SP treatment in NC and CAIII sh PC12 cells. Graphical representation of GO classification for biological processes, cellular components, and molecular functions (A,B) and GO enrichment analysis (C,D) for differential gene expression between DMSO and 60 μM SP groups in NC and CAIII sh cells, n = 3; KEGG pathway classification (E,F) and KEGG enrichment scatterplots (G,H) for the same comparison, n = 3. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Recombinant AAV vectors encoding CAIII shRNA (5′-GGTTCACTGGAATCCAAAGTA-3′) or non-specific control shRNA (5′-CGCTGAGTACTTCGAAATGTC-3′) were synthesized by Shanghai Genechem Co., Ltd. Mice were anesthetized with 1.2% isoflurane and placed on a stereotaxic apparatus (RWD, China) with a heating pad to maintain body temperature at 37°C.

Techniques: Gene Expression, Comparison