high glucose  (Thermo Fisher)

Journal: Redox Biology

Article Title: Astrocytic GSTM2-STAT3 interaction ameliorates the diabetes associated cognitive dysfunction via targeting mitochondrial defects and oxidative stress

doi: 10.1016/j.redox.2026.104137

Figure Lengend Snippet: Inhibition of STAT3/Drp1 signaling defends HG-induced oxidative stress and mitochondrial defects. (A) Western blotting of STAT3 protein levels in primary cultured astrocytes, and quantification data. n = 3. (B) Representative images of ROS content by flow cytometry analysis, and quantification data. n = 5. (C-E) MDA, GSH and SOD contents in primary cultured astrocytes. n = 5. (F) Representative images of the morphology of mitochondria by TEM in primary cultured astrocytes. Scale bar, 500 nm. (G) Mitochondrial aspect ratio, n = 3. (H) Mitochondrial cristae density, n = 3. (I) Western blotting of Mfn1, p -Drp1 and Drp1 protein levels in primary cultured astrocytes. (J) Representative images of JC-1 staining by flow cytometry analysis used to assess mitochondrial membrane potential. (K–N) OCR, ATP production, Basal respiration and Maximal respiration in primary cultured astrocytes. n = 5. si-NC (scrambled negative control siRNA); si-STAT3 (si-STAT3 primer). (High glucose, 50 mmol/L treated 48 h). Data are shown as mean ± SEM or SD. Statistical significance was defined as ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001.

Article Snippet: After reaching logarithmic growth, they were exposed to either high-glucose (HG) medium (50 mmol/L, A16828, Thermo Fisher) or mannitol (50 mmol/L, M8140-250, Beijing Solarbio Science & Technology Co., Ltd.) for 24, 48, or 72 h. Following treatment, 10 μL of CCK-8 solution was added to each well, and after a 1 h incubation at 37 °C, absorbance was measured to determine cell viability using a Cell Counting Kit-8 (CK04, Servicebio, Beijing, China).

Techniques: Inhibition, Western Blot, Cell Culture, Flow Cytometry, Staining, Membrane, Negative Control

Journal: Redox Biology

Article Title: Astrocytic GSTM2-STAT3 interaction ameliorates the diabetes associated cognitive dysfunction via targeting mitochondrial defects and oxidative stress

doi: 10.1016/j.redox.2026.104137

Figure Lengend Snippet: GSTM2 expression in astrocytes is significantly downregulated in the hippocampus of DACD mice models. (A) Experimental paradigms. (B) The represents of the exploration trajectory of db/m and db/db mice in the NOR test. (C) The preference index in the NOR test. n = 10. (D) The represents of the swimming trajectories in the MWM test. (E-G) The escape latency, across platform times and target quadrant retention time. n = 10. (H) Cluster analysis of proteomic sequencing. (I) Volcano plot volcanic map of proteomic sequencing. Red dots are raised, and green dots are decreased. Gstm2 was downregulated in hippocampus of db/db mice compared to db/m mice. (J) KEGG pathway enrichment analysis showed Glutathione metabolism was the key metabolic pathways. (K) Representative images of the cellular localization and expression of GSTM2 by immunofluorescence in astrocytes (GFAP), microglia (Iba-1), neurons (NeuN), oligodendrocytes (MBP) and nucleus (DAPI) of the CA1 region in hippocampus. Scale bar, 50 μm. And quantification data. n = 3. (L) Western blotting of GSTM2 protein expression in hippocampus, and quantification data. n = 3. (M) Representative images of the localization and expression of GSTM2 by immunofluorescence in HG primary cultured astrocytes (50 mmol/L concentration glucose treated for 48 h) compared to control. Scale bar, 50 μm. And quantification data. n = 3. (N) Western blotting of GSTM2 protein expression in primary cultured astrocytes, and quantification data. n = 3. HG (High glucose, 50 mmol/L treated 48 h). Data are shown as mean ± SEM. Statistical significance was defined as ∗∗ P < 0.01 and ∗∗ P < 0.001.

Article Snippet: After reaching logarithmic growth, they were exposed to either high-glucose (HG) medium (50 mmol/L, A16828, Thermo Fisher) or mannitol (50 mmol/L, M8140-250, Beijing Solarbio Science & Technology Co., Ltd.) for 24, 48, or 72 h. Following treatment, 10 μL of CCK-8 solution was added to each well, and after a 1 h incubation at 37 °C, absorbance was measured to determine cell viability using a Cell Counting Kit-8 (CK04, Servicebio, Beijing, China).

Techniques: Expressing, Sequencing, Immunofluorescence, Western Blot, Cell Culture, Concentration Assay, Control

Journal: Redox Biology

Article Title: Astrocytic GSTM2-STAT3 interaction ameliorates the diabetes associated cognitive dysfunction via targeting mitochondrial defects and oxidative stress

doi: 10.1016/j.redox.2026.104137

Figure Lengend Snippet: GSTM2 alleviates oxidative stress and mitochondrial defects in DACD models. (A) Representative images of the morphology of astrocytes mitochondria by TEM in hippocampus. Scale bar, 1.0 μm. (B) Mitochondrial aspect ratio, n = 3. (C) Mitochondrial cristae density, n = 3. (D-F) MDA, GSH and SOD contents in hippocampus. n = 5. (G) Western blotting of GSTM2 protein expression in primary cultured astrocytes, and quantification data. n = 3. (H) Representative images of ROS content by flow cytometry analysis, and quantification data. n = 5. (I–K) MDA, GSH and SOD contents in primary cultured astrocytes. n = 5. (L) Representative images of the morphology of primary astrocytes mitochondria by TEM. Scale bar, 500 nm. (M) Mitochondrial aspect ratio, n = 3. (N) Mitochondrial cristae density, n = 3. (O) Western blotting of Mfn1, p -Drp1 and Drp1 protein levels in primary cultured astrocytes, and quantification data. n = 3. (P, Q) Representative images of JC-1 staining by flow cytometry analysis used to assess mitochondrial membrane potential. (R–U) OCR, ATP production, Basal respiration and Maximal respiration in primary cultured astrocytes. n = 5. rAAV-NC (rAAV-GFAP-EGFP-WPRE-hGH pA); rAAV-GSTM2 (rAAV-GFAP-GSTM2-EGFP-WPRE-hGH pA). Vector (pEGFP-C1 plasmid empty vector); OE-GSTM2 (pEGFP-C1-GSTM2 plasmid vector). HG (High glucose, 50 mmol/L treated 48 h). Data are shown as mean ± SEM or SD. Statistical significance was defined as ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗ P < 0.001.

Article Snippet: After reaching logarithmic growth, they were exposed to either high-glucose (HG) medium (50 mmol/L, A16828, Thermo Fisher) or mannitol (50 mmol/L, M8140-250, Beijing Solarbio Science & Technology Co., Ltd.) for 24, 48, or 72 h. Following treatment, 10 μL of CCK-8 solution was added to each well, and after a 1 h incubation at 37 °C, absorbance was measured to determine cell viability using a Cell Counting Kit-8 (CK04, Servicebio, Beijing, China).

Techniques: Western Blot, Expressing, Cell Culture, Flow Cytometry, Staining, Membrane, Plasmid Preparation

Journal: Redox Biology

Article Title: Astrocytic GSTM2-STAT3 interaction ameliorates the diabetes associated cognitive dysfunction via targeting mitochondrial defects and oxidative stress

doi: 10.1016/j.redox.2026.104137

Figure Lengend Snippet: GSTM2 suppresses the phosphorylation of STAT3. (A) Experimental paradigms. (B) Top 15 list of GSTM2-binding proteins identified through GSTM2 IP-MS analysis in primary cultured astrocytes. STAT3 was identified with high level of interaction intensity. (C) Detection of the binding affinity between GSTM2 and STAT3 by SPR method. (D-E) Co-IP analysis for validation of the endogenous interaction between GSTM2 and STAT3 in primary cultured astrocytes. (F) Representative images of the cellular localization and expression of GSTM2 and STAT3 by immunofluorescence in primary cultured astrocytes. Scale bar, 50 μm. GSTM2 (red), STAT3 (green) and DAPI (blue). (G) Western blotting of p-STAT3(Ser727), STAT3 and GSTM2 protein levels in primary cultured astrocytes, and quantification data. n = 3. si-NC (scrambled negative control siRNA); si-GSTM2 (si-GSTM2 primer). Vector (pEGFP-C1 plasmid empty vector); OE-GSTM2 (pEGFP-C1-GSTM2 plasmid vector). HG (High glucose, 50 mmol/L treated 48 h). Data are shown as mean ± SEM. Statistical significance was defined as ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001.

Article Snippet: After reaching logarithmic growth, they were exposed to either high-glucose (HG) medium (50 mmol/L, A16828, Thermo Fisher) or mannitol (50 mmol/L, M8140-250, Beijing Solarbio Science & Technology Co., Ltd.) for 24, 48, or 72 h. Following treatment, 10 μL of CCK-8 solution was added to each well, and after a 1 h incubation at 37 °C, absorbance was measured to determine cell viability using a Cell Counting Kit-8 (CK04, Servicebio, Beijing, China).

Techniques: Phospho-proteomics, Binding Assay, Protein-Protein interactions, Cell Culture, Co-Immunoprecipitation Assay, Biomarker Discovery, Expressing, Immunofluorescence, Western Blot, Negative Control, Plasmid Preparation