Journal: Biomolecules

Article Title: A Critical Regulation of Th17 Cell Responses and Autoimmune Neuro-Inflammation by Ginsenoside Rg3

doi: 10.3390/biom10010122

Figure Lengend Snippet: Rg3 inhibited DC-mediated Th17 differentiation. ( A ) Experimental scheme. Naïve CD4 + T cells were incubated with LPS-stimulated BMDCs in the presence of soluble anti-CD3 and 1.5 ng/mL TGFβ. DMSO (vehicle) or indicated doses of Rg3 (18.75 or 37.5 μg/mL) was added at the beginning of cell culture. The frequency of IL-17A + CD4 + T cells and the level of IL-17A were determined by FACS ( B ) and ELISA ( C ), respectively. Data are mean ± SEM and represent three independent experiments. ** p < 0.01, *** p < 0.001 in comparison with the vehicle-treated group.

Article Snippet: After washing cells with cold PBS containing 1.5% FBS, cells were stained with APC-Cy7-conjugated anti-CD45.2 mAb and PE/Cy7-conjugated anti-CD4 mAb (eBioscience, San Diego, CA, USA) for surface staining.

Techniques: Incubation, Cell Culture, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Biomolecules

Article Title: A Critical Regulation of Th17 Cell Responses and Autoimmune Neuro-Inflammation by Ginsenoside Rg3

doi: 10.3390/biom10010122

Figure Lengend Snippet: Rg3 inhibited Th17 differentiation in a T-cell-intrinsic manner. ( A ) Experimental scheme. Naïve CD4 + T cells were stimulated with pre-coated anti-CD3 and CD28 in the presence of 10 ng/mL IL-6 and 1.5 ng/mL TGFβ. DMSO (vehicle) or indicated doses of Rg3 (18.75 or 37.5 μg/mL) was added at the beginning cell culture. The frequency of IL-17A + CD4 + T cells and the level of IL-17A were determined by FACS. ( B ) The amount of IL-17A and the transcript levels of indicated genes were measured by ELISA and quantitative RT-PCR, respectively ( C , D ). The concentration of Rg3 in ( C ) and ( D ) was 37.5 μg/mL. Data represent mean ± SEM and represent three independent experiments. *** p < 0.001 in comparison with the vehicle-treated group.

Article Snippet: After washing cells with cold PBS containing 1.5% FBS, cells were stained with APC-Cy7-conjugated anti-CD45.2 mAb and PE/Cy7-conjugated anti-CD4 mAb (eBioscience, San Diego, CA, USA) for surface staining.

Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Concentration Assay, Comparison

Journal: Biomolecules

Article Title: A Critical Regulation of Th17 Cell Responses and Autoimmune Neuro-Inflammation by Ginsenoside Rg3

doi: 10.3390/biom10010122

Figure Lengend Snippet: Rg3 diminished RORγt expression in T cells during Th17 differentiation. Naïve CD4 + T cells were stimulated with pre-coated anti-CD3 and CD28 in the presence of 1.5 ng/mL TGFβ and indicated concentrations of IL-6. DMSO (vehicle) or Rg3 (37.5 μg/mL) was added at the beginning of the cell culture. The frequency of IL-17A + ( A ) and IL-17A + RORγt + ( B ) CD4 + T cells was measured by FACS. ( C ) The histogram of RORγt + expression in CD4 + T cells is shown. Experiments were conducted three times. Data represent mean ± SEM. *** p < 0.001 in comparison with the vehicle-treated group.

Article Snippet: After washing cells with cold PBS containing 1.5% FBS, cells were stained with APC-Cy7-conjugated anti-CD45.2 mAb and PE/Cy7-conjugated anti-CD4 mAb (eBioscience, San Diego, CA, USA) for surface staining.

Techniques: Expressing, Cell Culture, Comparison

Journal: Biomolecules

Article Title: A Critical Regulation of Th17 Cell Responses and Autoimmune Neuro-Inflammation by Ginsenoside Rg3

doi: 10.3390/biom10010122

Figure Lengend Snippet: Rg3 inhibited the expansion of myelin oligodendrocyte glycoprotein (MOG)-reactive Th17 cells. ( A ) Experimental scheme. Mice were immunized s.c. with MOG in CFA. Seven days later, the lymphoid cells from the draining lymph nodes (LNs) were harvested and restimulated with MOG and IL-23 in the presence of DMSO (vehicle) or Rg3 (37.5 μg/mL) for 5 days. ( B ) The expression of IL-17A and IFNγ in CD4 + T cells were examined by FACS. Data shown are mean ± SEM and represent one from two independent experiments. ** p < 0.01, *** p < 0.001 in comparison with the vehicle-treated group.

Article Snippet: After washing cells with cold PBS containing 1.5% FBS, cells were stained with APC-Cy7-conjugated anti-CD45.2 mAb and PE/Cy7-conjugated anti-CD4 mAb (eBioscience, San Diego, CA, USA) for surface staining.

Techniques: Expressing, Comparison

Journal: Biomolecules

Article Title: A Critical Regulation of Th17 Cell Responses and Autoimmune Neuro-Inflammation by Ginsenoside Rg3

doi: 10.3390/biom10010122

Figure Lengend Snippet: Rg3 decreases autoimmune neuro-inflammation induced by adoptive transfer of MOG-reactive CD4 + T cells. ( A ) Experimental scheme of passive experimental autoimmune encephalomyelitis (EAE), induction. ( B – D ) EAE incidence, % weight change, and clinical score were monitored daily. The levels of IL-17A and IFNγ in CD4 + T cells were examined in the CNS ( E ) and the inguinal LN ( F ) on day 13 after EAE induction. Data shown are mean ± SEM and represent two independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 in comparison with the vehicle-treated group.

Article Snippet: After washing cells with cold PBS containing 1.5% FBS, cells were stained with APC-Cy7-conjugated anti-CD45.2 mAb and PE/Cy7-conjugated anti-CD4 mAb (eBioscience, San Diego, CA, USA) for surface staining.

Techniques: Adoptive Transfer Assay, Comparison