Journal: Oncotarget

Article Title: Immethridine, histamine H 3 -receptor (H 3 R) agonist, alleviated experimental autoimmune encephalomyelitis via inhibiting the function of dendritic cells

doi: 10.18632/oncotarget.20500

Figure Lengend Snippet: (A) FACS analysis of CD4 + T cell subsets in splenocytes of EAE mice (n=6). Splenocytes were prepared and Th1 and Th17 cells were analyzed by intracellular staining of IFN-γ and IL-17A, respectively, in the CD4 + gate. *p<0.05, **p<0.01 versus vehicle. (B) MTT analysis of MOG 35-55 -specific T cell proliferation. Splenocytes of EAE mice (n=6) were isolated and cultured at the present or absent of MOG 35-55 peptides for 72h and T cell proliferation was measured using a MTT Cell Proliferation Assay Kit. The data were shown at stimulation index. **p<0.01 versus vehicle. (C) FACS analysis of expression of co-stimulatory molecules on DCs in splenocytes of EAE mice (n=6). Splenocytes were prepared and stained with fluorescent labeled-antibodies against CD11c, CD40 or CD86, the stained cells were analyzed on flow cytometry. *p<0.05, **p<0.01 versus vehicle. (D) FACS analysis of expression of MHCII on DCs in spinal cord of EAE mice (n=6). Immune cells in spinal cord were prepared and stained with fluorescent labeled-antibodies against CD11c, MHCII, the stained cells were analyzed on flow cytometry. ***p<0.001 versus vehicle. (E) FACS analysis of CD4 + T cell subsets in spinal cord of EAE mice (n=6). The lymphocytes from spinal cord were prepared and Th1 and Th17 cells were analyzed by intracellular staining of IFN-γ and IL-17A, respectively, in the CD4 + gate. *p<0.05 versus vehicle. Data are representative of three independent experiments.

Article Snippet: The plates were incubated at 37 °C in a humid atmosphere with 5% CO 2 for 5 days and the result was measured by MTT Cell Proliferation Kit (Beyotime, China) according to the manufacturer’s instructions.

Techniques: Staining, Isolation, Cell Culture, MTT Cell Proliferation, Expressing, Labeling, Flow Cytometry

Journal: Oncotarget

Article Title: Immethridine, histamine H 3 -receptor (H 3 R) agonist, alleviated experimental autoimmune encephalomyelitis via inhibiting the function of dendritic cells

doi: 10.18632/oncotarget.20500

Figure Lengend Snippet: FACS analysis of CD40 and CD86 on BMDCs. Immature DCs were exposed to combinations of immethridine and/or LPS for 12 h, and cells were stained with a FITC-labeled anti-CD11c and PE-labeled anti-CD40 (A) or anti-CD86 (B) , then the results were analyzed by FACS. **p<0.01, ***p<0.001 versus PBS. Data are representative of three experiments. (C) Immethridine inhibited the capacity of antigen-presentation of DCs. MOG-pulsed DCs was co-cultured with MOG-specific CD4 + T for 72h and T cell proliferation was detected by MTT Cell Proliferation Assay Kit. The data were shown in relative T cell proliferation. *p<0.05 versus PBS. (D) MLR assay. Purified T cells from BALB/C mice and immethridine-treated or untreated DCs were co-cultured for 5 days and T cell proliferation was detected by MTT Cell Proliferation Assay Kit. The data was shown in OD value at 570nm. *p<0.05 versus PBS. Data are representative of three experiments.

Article Snippet: The plates were incubated at 37 °C in a humid atmosphere with 5% CO 2 for 5 days and the result was measured by MTT Cell Proliferation Kit (Beyotime, China) according to the manufacturer’s instructions.

Techniques: Staining, Labeling, Immunopeptidomics, Cell Culture, MTT Cell Proliferation, Mlr Assay, Purification