Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD314 , REA1228 , 50 , 130-124-341 , PE , Miltenyi Biotec.

Techniques: Imaging

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: Cancers

Article Title: Multiparametric Characterization of the DSL-6A/C1 Pancreatic Cancer Model in Rats

doi: 10.3390/cancers16081535

Figure Lengend Snippet: List of antibodies used for the immunological analysis of the DSL-6A/C1 tumors.

Article Snippet: CD314 (NKG2D) , APC , REA471 , Human cell line , Miltenyi , 130-106-992.

Techniques: