Journal: Neurochemical Research

Article Title: Morin Improves Cognitive Deficits in an in Vivo Model of Vascular Dementia by Modulating the N-methyl-D-aspartate Receptor Signaling Pathways

doi: 10.1007/s11064-026-04717-7

Figure Lengend Snippet: Morin modulated the expression of NMDA receptors in the hippocampus of VaD rats. a the expression levels of NR1 ; b the expression levels of NR2A ; c the expression levels of NR2B ; d the expression levels of NR1 protein; e the expression levels of NR2A protein; f the expression levels of NR2B protein; g protein levels of p-CREB; h protein levels of p-CAMK2A; i protein levels of p-CAMK2D. Protein levels of NR1, NR2A, and NR2B were quantified by ELISA. Data are presented as mean ± SD ( n = 8 per group). Statistical analysis was performed by one-way ANOVA with Tukey’s post-hoc test (data met assumptions of normality and homoscedasticity)/Kruskal-Wallis with Dunn’s test (data did not meet assumptions). * indicates a significant difference from the Sham group; # indicates a significant difference 2VO group; *# indicates a significant difference from both the Sham and 2VO groups, with a p-value of less than 0.05 considered statistically significant

Article Snippet: Moreover, phosphorylation levels of calcium/calmodulin-dependent protein kinase II isoforms CAMK2A and CAMK2D at Thr286 (p-CAMK2A, p-CAMK2D) were quantified using the Colorimetric Cell-Based ELISA Kit (CAMK2A/CAMK2D (Phospho-Thr286), Boster Bio, #EKC2366).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: Science immunology

Article Title: The human CD47 checkpoint is targeted by an immunosuppressive Aedes aegypti salivary factor to enhance arboviral skin infectivity

doi: 10.1126/sciimmunol.adk9872

Figure Lengend Snippet: ( A and B ) Blood-derived human primary macrophages and macrophage-like THP-1 and U937 human cell lines were used in these experiments. Phagocytosis activity was measured 4 hours after stimulation (A). Phagocytosis assay with Nest1 and GlycA-preincubated RBCs (B). Blue bars, no treatment; yellow bars, Nest1 treatment; purple bars, GlycA); orange bars, Nest1+GlycA. These values represent the means ± SEM of three replicates from a single experiment. The P value is displayed in the graph and was determined using Student’s t test. Experiments with THP-1 and human primary macrophages are representative of two or three independent experiments. ( C ) Relative expression of proinflammatory cytokines (IFN-γ, IL-2, TNF, IL-12p40, IL-8, and IL-6) and anti-inflammatory cytokines (IL-13, IL-4, IL-5, and IL-1RA) from human white blood cells from four different donors. Expression was measured by qRT-PCR in nontreated and Nest1-treated human white blood cells 24 hours after stimulation. Cells were stimulated in triplicate, and the measurements of four different donors were pooled. Human GAPDH was used as a housekeeping gene. These values represent the means ± SEM of three replicates from a single experiment. Asterisks represent significant difference between samples, calculated by Student’s t test (* P < 0.05, ** P < 0.01, and *** P < 0.001).

Article Snippet: Cells were incubated for 4 hours at 37°C in the presence or absence of Nest1 protein and anti-human GlycA (BioxCell InVivoMAb, 6A7M) antibodies as opsonin at different concentrations.

Techniques: Derivative Assay, Activity Assay, Phagocytosis Assay, Expressing, Quantitative RT-PCR

Journal: American Journal of Cancer Research

Article Title: TGF-β1 expression in regulatory NK1.1 - CD4 + NKG2D + T cells dependents on the PI3K-p85α/JNK, NF-κB and STAT3 pathways

doi:

Figure Lengend Snippet: TGF-β1 production of NK1.1-CD4+NKG2D+ cells upon various stimulations at different times. A. TGF-β1 of NK1.1-CD4+NKG2D+ cells detected by flow cytometry at 0.5-, 2-, 8-, 16-hour stimulation. B. Statistical analysis of TGF-β1 variations at different time points. C. TGF-β1 transcription of NK1.1-CD4+NKG2D+ cells detected by real-time PCR. D. Concentration of TGF-β1 in supernatants of NK1.1-CD4+NKG2D+ cells upon indicated stimulations measured by ELISA. Each experiment was repeated at least thrice. *, P<0.05; **, P<0.01; ***, P<0.001; ns, no significance.

Article Snippet: Splenic NK1.1 - CD4 + T cells were enriched from splenic single cell suspension by mouse CD4 + T lymphocyte enrichment set (BD Biosciences) following the manufacturer’s instructions, and then NKG2D + T cells were isolated by indirectly labeled the cells with PE-conjugated anti-mouse NKG2D antibody (CX5) and anti-PE MicroBeads (Miltenyi Biotec GmbH; Bergisch Gladbach, Germany).

Techniques: Flow Cytometry, Real-time Polymerase Chain Reaction, Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: American Journal of Cancer Research

Article Title: TGF-β1 expression in regulatory NK1.1 - CD4 + NKG2D + T cells dependents on the PI3K-p85α/JNK, NF-κB and STAT3 pathways

doi:

Figure Lengend Snippet: Activation of PI3K, MAPK, STAT3 and NF-κB signaling pathways. Western blot analysis of PI3K-p85, -p110, Akt, pAkt in NK1.1-CD4+NKG2D+ cells upon different stimulations at 2-hour (A) or 8-hour (C). Statistical analysis of expression variations of PI3K-p85, -p110, Akt, pAkt at 2-hour (B) or 8-hour (D). Analysis of JNK, Erk, p38 (E, F), NF-κB, STAT3 (G, H) and their phosphorylation in NK1.1-CD4+NKG2D+ cells upon stimulations for 8 hours. Each experiment was repeated at least thrice. *, P<0.05; **, P<0.01; ***, P<0.001; ns, no significance.

Article Snippet: Splenic NK1.1 - CD4 + T cells were enriched from splenic single cell suspension by mouse CD4 + T lymphocyte enrichment set (BD Biosciences) following the manufacturer’s instructions, and then NKG2D + T cells were isolated by indirectly labeled the cells with PE-conjugated anti-mouse NKG2D antibody (CX5) and anti-PE MicroBeads (Miltenyi Biotec GmbH; Bergisch Gladbach, Germany).

Techniques: Activation Assay, Protein-Protein interactions, Western Blot, Expressing, Phospho-proteomics

Journal: American Journal of Cancer Research

Article Title: TGF-β1 expression in regulatory NK1.1 - CD4 + NKG2D + T cells dependents on the PI3K-p85α/JNK, NF-κB and STAT3 pathways

doi:

Figure Lengend Snippet: Inhibition of PI3K/JNK/AP-1 on TGF-β1 expression. NK1.1-CD4+NKG2D+ cells were treated with the PI3K inhibitor (LY294002) under different stimulations for 8 hours (A). (B) Statistical analysis of effects of LY294002 on TGF-β1 expression. (C, D) Effects on JNK/pJNK expression by the PI3K inhibitor (LY294002). Effects of JNK inhibitor (E, F), Erk or p38 inhibitor (G), and AP-1 inhibitor (H, I) on TGF-β1 expression of NK1.1-CD4+NKG2D+ cells. Each experiment was repeated at least thrice. *, P<0.05; **, P<0.01; ***, P<0.001; ns, no significance.

Article Snippet: Splenic NK1.1 - CD4 + T cells were enriched from splenic single cell suspension by mouse CD4 + T lymphocyte enrichment set (BD Biosciences) following the manufacturer’s instructions, and then NKG2D + T cells were isolated by indirectly labeled the cells with PE-conjugated anti-mouse NKG2D antibody (CX5) and anti-PE MicroBeads (Miltenyi Biotec GmbH; Bergisch Gladbach, Germany).

Techniques: Inhibition, Expressing

Journal: American Journal of Cancer Research

Article Title: TGF-β1 expression in regulatory NK1.1 - CD4 + NKG2D + T cells dependents on the PI3K-p85α/JNK, NF-κB and STAT3 pathways

doi:

Figure Lengend Snippet: Inhibition of NF-κB on TGF-β1 expression. Effects of Bortezomib (NF-κB inhibitor) on TGF-β1 expression (A, B). Effects of LY294002 on NF-κB p65 activation in NK1.1-CD4+NKG2D+ cells for 8 hours (C, D). Each experiment was repeated at least thrice. *, P<0.05; **, P<0.01; ***, P<0.001; ns, no significance.

Article Snippet: Splenic NK1.1 - CD4 + T cells were enriched from splenic single cell suspension by mouse CD4 + T lymphocyte enrichment set (BD Biosciences) following the manufacturer’s instructions, and then NKG2D + T cells were isolated by indirectly labeled the cells with PE-conjugated anti-mouse NKG2D antibody (CX5) and anti-PE MicroBeads (Miltenyi Biotec GmbH; Bergisch Gladbach, Germany).

Techniques: Inhibition, Expressing, Activation Assay

Journal: American Journal of Cancer Research

Article Title: TGF-β1 expression in regulatory NK1.1 - CD4 + NKG2D + T cells dependents on the PI3K-p85α/JNK, NF-κB and STAT3 pathways

doi:

Figure Lengend Snippet: Inhibition of STAT3 on TGF-β1 expression. Effects of STAT3 inhibitors on TGF-β1 expression in NK1.1-CD4+NKG2D+ cells for 8 hours (A). (B, C) Effects of LY294002 on STAT3-pY705 phosphorylation. (D, E) Effects of Bortezomib on STAT3-pY705 phosphorylation. (F) The predicted binding sites of the TGF-β promoter by STAT3. (G) STAT3 engagement measured by a ChIP assay. Each experiment was repeated at least thrice. *, P<0.05; **, P<0.01; ***, P<0.001; ns, no significance.

Article Snippet: Splenic NK1.1 - CD4 + T cells were enriched from splenic single cell suspension by mouse CD4 + T lymphocyte enrichment set (BD Biosciences) following the manufacturer’s instructions, and then NKG2D + T cells were isolated by indirectly labeled the cells with PE-conjugated anti-mouse NKG2D antibody (CX5) and anti-PE MicroBeads (Miltenyi Biotec GmbH; Bergisch Gladbach, Germany).

Techniques: Inhibition, Expressing, Phospho-proteomics, Binding Assay

Journal: American Journal of Cancer Research

Article Title: TGF-β1 expression in regulatory NK1.1 - CD4 + NKG2D + T cells dependents on the PI3K-p85α/JNK, NF-κB and STAT3 pathways

doi:

Figure Lengend Snippet: Diagram of the TGF-β1 transcription in NK1.1-CD4+NKG2D+ cells regulated by AP-1, NF-κB, and STAT3.

Article Snippet: Splenic NK1.1 - CD4 + T cells were enriched from splenic single cell suspension by mouse CD4 + T lymphocyte enrichment set (BD Biosciences) following the manufacturer’s instructions, and then NKG2D + T cells were isolated by indirectly labeled the cells with PE-conjugated anti-mouse NKG2D antibody (CX5) and anti-PE MicroBeads (Miltenyi Biotec GmbH; Bergisch Gladbach, Germany).

Techniques:

Journal: Neurochemical Research

Article Title: Morin Improves Cognitive Deficits in an in Vivo Model of Vascular Dementia by Modulating the N-methyl-D-aspartate Receptor Signaling Pathways

doi: 10.1007/s11064-026-04717-7

Figure Lengend Snippet: Morin modulated the expression of NMDA receptors in the hippocampus of VaD rats. a the expression levels of NR1 ; b the expression levels of NR2A ; c the expression levels of NR2B ; d the expression levels of NR1 protein; e the expression levels of NR2A protein; f the expression levels of NR2B protein; g protein levels of p-CREB; h protein levels of p-CAMK2A; i protein levels of p-CAMK2D. Protein levels of NR1, NR2A, and NR2B were quantified by ELISA. Data are presented as mean ± SD ( n = 8 per group). Statistical analysis was performed by one-way ANOVA with Tukey’s post-hoc test (data met assumptions of normality and homoscedasticity)/Kruskal-Wallis with Dunn’s test (data did not meet assumptions). * indicates a significant difference from the Sham group; # indicates a significant difference 2VO group; *# indicates a significant difference from both the Sham and 2VO groups, with a p-value of less than 0.05 considered statistically significant

Article Snippet: Moreover, phosphorylation levels of calcium/calmodulin-dependent protein kinase II isoforms CAMK2A and CAMK2D at Thr286 (p-CAMK2A, p-CAMK2D) were quantified using the Colorimetric Cell-Based ELISA Kit (CAMK2A/CAMK2D (Phospho-Thr286), Boster Bio, #EKC2366).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay

Journal: The journal of medical investigation : JMI

Article Title: Antifibrotic effects of CXCR4 antagonist in bleomycin-induced pulmonary fibrosis in mice.

doi: 10.2152/jmi.60.127

Figure Lengend Snippet: Figure 6 Administration of AMD3100 reduces the number of fibrocytes into the lungs treated with bleomycin in mice. Mice were implanted with osmotic minipumps containing bleomycin (BLM). AMD3100 (6 mg/kg/day) was injected using osmotic minipump. Single-cell suspensions were isolated from the lungs of bleomycin-treated mice on day 7. After culturing in fibronectin- coated 10 cm-dish, the adherent cells were stained for CD45 and Collagen I, and examined by flow cytometry. A : Flow cytometric analysis of fibrocytes. The data shows the double staining with anti-CD45 and anti-collagen I antibodies for adherent cells of the lungs of bleomycin-treated mice. B : The percentage of CD45+Collagen I+ fibrocytes in bleomycin-treated lungs. Data are shown as the meanSD of 4 mice. *p0.05, significant differ- ences between bleomycin alone and bleomycin+AMD3100 (6 mg/kg/day). Figure 5 AMD3100 inhibits migration of human fibrocytes in response to CXCL12. A : Generation of human fibrocytes. Hu- man fibrocytes were isolated from human peripheral blood mono- nuclear cells as described in the Methods. Cell surface markers of human fibrocytes were confirmed by flow cytometry. Human fibrocytes were stained with control antibody (closed black) and specific antibodies (open gray) for collagen I, CXCR4 and CD45. B : Migration of human fibrocytes in response to CXCL12 and its inhibition by AMD3100. Fibrocytes(1104/well) were added in upper chamber with or without AMD3100 (1 μg/ml). Fibrocytes were tested for chemotaxis (n=3) to CXCL12 (10 and 100 ng/ml). Data are shown as the meanSD of three wells. Similar results were obtained in four separate experiments. *p0.001 versus CXCL12 (100 ng/ml) alone.

Article Snippet: PE - conjugated anti - mouse CXCR4 antibody, PE-Cy5 conjugated anti-mouse CD45 antibody (BD Biosciences-Pharmingen, San Diego, California, USA), anti-collagen type I (Rockland, Gilbertsville, Pennsylvania, USA), BD Cytofix/ CytoPerm Plus Kit (BD Biosciences-Pharmingen), Collagenase IV (Gibco 17104-019), DEOXYRIBONUCLEASE (DNase) I (CALBIOCHEM, Madison, WI, USA) was used in this study.

Techniques: Injection, Isolation, Staining, Flow Cytometry, Double Staining, Migration, Control, Inhibition, Chemotaxis Assay