Journal: Experimental and Therapeutic Medicine

Article Title: Application of red clover isoflavone extract as an adjuvant in mice

doi: 10.3892/etm.2019.8315

Figure Lengend Snippet: Effect of RCIE or alum adjuvant on CD4 + and CD8 + T cell populations in the peripheral blood of OVA-immunized mice. Representative flow cytometry dot plots showing the population of CD4 + and CD8 + T cells in the peripheral blood from mice in the (A) Saline, (B) OVA, (C) OVA + alum, (D) OVA + 50 µg RCIE, (E) OVA + 100 µg RCIE and (F) OVA + 200 µg RCIE groups. (G) Quantified flow cytometry data. Values are presented as mean ± SEM (n=10). *P<0.05 vs. OVA. OVA, ovalbumin; alum, aluminum hydroxide gel; RCIE, red clover isoflavone extract; PE, phycoerythrin.

Article Snippet: The ELISA kits (IFN-γ, cat. no. F10660; TNF-α cat. no. F11630; IL-2 cat. no. F10780; IL-4 cat. no. F10810; and IL-5 cat. no. F10820) were from Shanghai Westang Biotechnology Co., Ltd. FITC/phycoerythrin (PE) rat anti-mouse CD4/CD8 antibodies (cat. no. FMD001-050) were purchased from Beijing 4A Biotech Co., Ltd. RNAiso plus ® (cat. no. 9108), First Strand cDNA Synthesis kit (cat. no. RR036A) and SYBR ® Premix Ex Taq™ were purchased from Takara Biotechnology Co., Ltd. Pathogenic porcine Escherichia coli ( E. coli ) ( ) was kindly provided by Dr Jingxuan Ni (Longyan University).

Techniques: Flow Cytometry

Journal: Cellular and Molecular Life Sciences

Article Title: Regulatory role and mechanisms of myeloid TLR4 in anti-GBM glomerulonephritis

doi: 10.1007/s00018-021-03936-1

Figure Lengend Snippet: Deficiency of myeloid TLR4 promotes macrophage polarization from M1 towards M2, blunts macrophage MHC II expression and further reduces T cell accumulation in experimental anti-GBM crescentic GN. Representative flow cytometry plots and quantification of renal singlets analyzed for A M1 (F4/80 + iNOS + ), mean ± SEM ( t test, n = 6–7) and B M2 (F4/80 + CD206 + ), mean ± SEM ( t test, n = 7–8) (both gated on CD45 + cells). Realtime PCR analysis of C il-1β, mean ± SEM (ANOVA, n = 6–10) and D il-10, mean ± SEM (ANOVA, n = 6–10) in kidneys isolated from control or diseased mice on day7 and 14 after anti-GBM GN induction. E Representative flow cytometry plots and quantification of renal singlets analyzed for F4/80 + MHC II + cells. Gated on CD45 + cells, mean ± SEM ( t test, n = 4–6). F Representative kidney sections stained for CD3 + (magnification × 40) and CD4 + T cells, (magnification × 100). G Representative flow cytometry plots and quantification of renal singlets analyzed for CD3 + , mean ± SEM ( t test, n = 4–7) and CD4 + T cells, mean ± SEM ( t test, n = 5–8) (Gated on leukocyte in FSC vs. SSC plot). Each dot represents one mouse. * p < 0.05, *** p < 0.001 versus corresponding control; # p < 0.05, ## p < 0.01 versus corresponding tlr4 f/f

Article Snippet: CD4 cells infiltrating in the kidney were identified in snap-frozen sections (4 μm) by immunofluorescence with Dylight 550-rat anti-mouse CD4 monoclonal antibody (Leinco Technologies, St. Louis, Missouri, USA).

Techniques: Expressing, Flow Cytometry, Isolation, Control, Staining

Journal: Cellular and Molecular Life Sciences

Article Title: Regulatory role and mechanisms of myeloid TLR4 in anti-GBM glomerulonephritis

doi: 10.1007/s00018-021-03936-1

Figure Lengend Snippet: Deficiency of myeloid TLR4 suppresses T cell-mediated experimental anti-GBM crescentic GN by shifting the Th1/Th17 towards the Treg immune responses. Representative flow cytometry plots of renal singlets analyzed for: A Th1 (CD4 + IFNγ + ), mean ± SEM ( t test, n = 5–8); B Th17 (CD4 + IL17a + ), mean ± SEM ( t -test, n = 5–8); C. Treg (CD4 + CD25 + Foxp3 + ), mean ± SEM ( t test, n = 6–8); and D Th2 (CD4 + IL-4 + ), mean ± SEM ( t test, n = 3–6) (Gated on CD4 + cells for Treg, otherwise gated on CD45 + ). Each dot represents one mouse. # p < 0.05, ## p < 0.01, ### p < 0.001 versus corresponding tlr4 f/f

Article Snippet: CD4 cells infiltrating in the kidney were identified in snap-frozen sections (4 μm) by immunofluorescence with Dylight 550-rat anti-mouse CD4 monoclonal antibody (Leinco Technologies, St. Louis, Missouri, USA).

Techniques: Flow Cytometry

Journal:

Article Title: Effect of an inhibitor of Jun N-terminal protein kinase, SP600125, in single allergen challenge in sensitized rats

doi: 10.1111/j.1365-2567.2004.01887.x

Figure Lengend Snippet: Effect of SP600125 on eosinophil and T-lymphocyte subset (CD2+, CD4+ and CD8+) counts in airway submucosa per mm of basement membrane for groups as detailed in Fig. 1. Allergen challenge caused a significant increase in T lymphocytes expressing CD2+. (*P < 0·05 as compared to saline group), and in MBP + eosinophils (***P < 0·001 as compared to the saline group). Data are shown as mean ± SEM.

Article Snippet: For staining of CD2 + , CD4 + , CD8 + T lymphocytes in tissue sections, sections were incubated with either mouse anti-rat CD2, CD4, or CD8 monoclonal antibody (pan T-cell markers, Pharmingen, Cambridge Bioscience, Cambridge, UK).

Techniques: Membrane, Expressing, Saline