Expressing:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
Control:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
Quantitative RT-PCR:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
Derivative Assay:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
In Vitro:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
Reverse Transcription Polymerase Chain Reaction:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
Gene Expression:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
Two Tailed Test:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
Activation Assay:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
In Vivo:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
Labeling:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
Binding Assay:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
Inhibition:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
Western Blot:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
Irradiation:Article Title: The MicroRNA miR-22 Represses Th17 Cell Pathogenicity by Targeting PTEN-Regulated Pathways.
Article Snippet: 16-0038-85, 5mg/ml; Thermo Fisher Scientific) and anti-CD28 mAb (catalog no. 16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).16- 0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral conditions (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific] and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]). ... The following conditions were used: Th17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1 [R&D Systems]; catalog no. 406-ML-025, 20 ng/ml IL-6 [R&DSystems]; catalog no. 16-7041- 95, 10mg/ml anti–IL-4mAb [ThermoFisher]; and catalog no. 16- 7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), Th17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6 [R&D Systems]; catalog no. 401-ML-025, 20 ng/ml IL-1b [R&D Systems]; catalogno.. 1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems] and catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]), Treg condition (catalog no. 402-ML-020, 5 ng/ml IL-2 [R&DSystems]; catalog no. 7666-MB-005, 2.5 ng/ml TGF-b1 [R&D Systems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]), and Th0 condition (catalog no. 16- 7041-95, 10mg/ml anti–IL-4mAb [Thermo Fisher Scientific] and catalog no. 16-7311-38, 10mg/ml anti–IFN-gmAb [Thermo Fisher Scientific]).1887-ML-010, 20ng/ml IL-23 [R&DSystems]; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb [Thermo Fisher Scientific]; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb [Thermo Fisher Scientific]), Th1 condition (catalog no. 419-ML010, 10 ng/ml IL-12 [R&D Systems]..
Article Title: Inhibition of Glycolysis in Pathogenic T H 17 Cells through Targeting a miR -21-Peli1 -c-Rel Pathway Prevents Autoimmunity.
Article Snippet: Naive CD4+ T cells were isolated from lymph nodes or spleen of 6–8-wk-old mice using a Naive CD4+ T Cell Isolation Kit (catalog no. 130-104-453; Miltenyi Biotec) according to the manufacturer’s instruction.. Naive CD4+ T cells were stimulated with plate-bound anti-CD3 mAb (catalog no. 16-0038-85, 5 mg/ml; Thermo Fisher Scientific) in the presence of antiCD28 mAb (catalog no. 16-0289-85, 2 mg/ml; Thermo Fisher Scientific) in a 48-well plate under neutral condition (catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), TH17 (b) condition (catalog no. 7666-MB-005, 2 ng/ml TGF-b1; R&D Systems; catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific), and TH17 (23) condition (catalog no. 406-ML-025, 20 ng/ml IL-6; R&D Systems; catalog no. 401-ML-025, 20 ng/ml IL-1b; R&D Systems; catalog no. 1887-ML-010, 20 ng/ml IL-23; R&D Systems; catalog no. 16-7041-95, 10 mg/ml anti–IL-4 mAb; Thermo Fisher Scientific; and catalog no. 16-7311-38, 10 mg/ml anti–IFN-g mAb; Thermo Fisher Scientific).. Cells were cultured in complete IMDM medium (Life Technology) supplemented with 10% FBS and 2-ME (1:1000).
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