Journal: Frontiers in Immunology

Article Title: Dim Light at Night Impairs Daily Variation of Circulating Immune Cells and Renal Immune Homeostasis

doi: 10.3389/fimmu.2020.614960

Figure Lengend Snippet: Dim light at night (dLAN) disturbs the daily variation and alters the number of circulating immune cells. (A–C) Flow cytometric analysis of peripheral white blood cells (WBCs) collected at ZT9 and ZT21 from rats exposed to either the control light–dark (LD) regime (CTRL) or dLAN (~2 lx) for 2 (upper rows) and 5 (lower rows) weeks. Data represent the mean ± SEM with n = 6–9 per group. (B) Numbers of CD4 + helper (Th) and CD8 + cytotoxic (Tc) T cells and the CD4/CD8 ratio. (C) Numbers of classical (CD43 lo HIS48 hi ) and non-classical (CD43 lo HIS48 hi ) monocytes. Significant differences were evaluated by two-way repeated ANOVA with the Tukey post hoc test for multiple comparisons. Only significant main effects (ZT and dLAN) or interactions (ZT×dLAN) are shown. Dotted lines indicate significant differences between individual groups if an interaction was significant. # P < 0.1, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: The following fluorochrome-conjugated monoclonal antibodies were used: PE-Cy7 anti-rat CD45 (clone OX-1; Sony Biotechnology, USA; 1611065), FITC anti-rat CD3 (clone G4.18; Thermo Fisher Scientific, USA; 11-0030), APC anti-rat CD4 (clone OX-35; Thermo Fisher Scientific; 17-0040), PE anti-rat CD8a (clone OX-8; Thermo Fisher Scientific; 12-0084), PE anti-rat CD45RA (clone OX-33; Thermo Fisher Scientific; MR6404), APC anti-rat CD161a (clone 10/78; Thermo Fisher Scientific; MR6805), FITC anti-rat granulocytes (clone HIS48; Thermo Fisher Scientific; 11-0570), and PE anti-rat CD43 (clone W3/13; Sony Biotechnology; 1614060).

Techniques: Control

Journal: Journal of Neuroinflammation

Article Title: Perturbing chondroitin sulfate proteoglycan signaling through LAR and PTPσ receptors promotes a beneficial inflammatory response following spinal cord injury

doi: 10.1186/s12974-018-1128-2

Figure Lengend Snippet: List of antibodies used in this study

Article Snippet: CD4 , Cedarlane (Sheep, AF6439) , WB , 1:100.

Techniques: Blocking Assay

Journal: Journal of Neuroinflammation

Article Title: Perturbing chondroitin sulfate proteoglycan signaling through LAR and PTPσ receptors promotes a beneficial inflammatory response following spinal cord injury

doi: 10.1186/s12974-018-1128-2

Figure Lengend Snippet: List of flow cytometry antibodies used in this study

Article Snippet: CD4 , Cedarlane (Sheep, AF6439) , WB , 1:100.

Techniques: Flow Cytometry, Control

Journal: Journal of Neuroinflammation

Article Title: Perturbing chondroitin sulfate proteoglycan signaling through LAR and PTPσ receptors promotes a beneficial inflammatory response following spinal cord injury

doi: 10.1186/s12974-018-1128-2

Figure Lengend Snippet: ILP and ISP promotes a phenotypic switch in helper T cells toward a T reg phenotype. a , b Flow cytometric assessment revealed no apparent difference in the overall infiltration of helper T cells (CD45 + /CD3 + /CD4 + ) in the injured spinal cord at 3 and 7 days post-injury between vehicle and ILP/ISP-treated animals. c , d However, ILP/ISP-treated animals exhibited a significant decrease in the number of effector T cells (CD45 + /CD3 + /CD4 + /IFNγ + ) at 7 days post-injury. e , f A significant increase in the total number of regulatory T cells (CD45 + /CD3 + /CD4 + /IL10 + ) was observed at 3 days post-injury in ILP/ISP-treated animals. g , h Representative flow cytometry gates are shown. i Western blot analysis showed upregulation of CD4 protein expression, at 7 and 14 days post-SCI compared to uninjured control group confirming infiltration of helper T cells in the injured spinal cord. Confirming our flow cytometry, ILP and ISP had no apparent effect on the overall protein expression of CD4. j However, ILP and ISP significantly increased FOXP3 protein expression, a marker of regulatory T cells, at both 7 and 14 days post-SCI compared to SCI vehicle control. Western blot results have been normalized to the actin loading control prior to subsequent normalization to the control values. The data show mean ± SEM, * p < 0.05, Student t test ( a – f ), one-way ANOVA ( i , j ), N = 4-6/group

Article Snippet: CD4 , Cedarlane (Sheep, AF6439) , WB , 1:100.

Techniques: Flow Cytometry, Western Blot, Expressing, Control, Marker

Journal: Physiological Genomics

Article Title: Mycophenolate mofetil prevents cerebrovascular injury in stroke-prone spontaneously hypertensive rats

doi: 10.1152/physiolgenomics.00110.2016

Figure Lengend Snippet: Renal immune cell infiltration. Immunohistochemistry was used to detect CD43+ lymphocytes (top) and CD68+ macrophages (bottom) infiltration in the kidneys collected from vehicle-treated (−MMF) or MMF-treated (+MMF) rats at the end of the treatment protocol or at the time of euthanasia. Representative images are shown above. Scale bar = 50 μm for CD43 and 20 μm for CD68 images. A large number of infiltrating CD43+ and CD68+ cells (indicated by arrows) were detected in the kidneys of vehicle-treated SHR-A3 rats. Immune cell infiltration was markedly reduced by immunosuppression with MMF for 8 wk; n = 8–9. *P < 0.05 vs. vehicle-treated (−MMF) rats.

Article Snippet: CD68 (clone ED1, ABD Serotec) was used as a marker of activated microglia/macrophages, and CD43 (clone W3/13, ThermoFisher Scientific) was used to identify lymphocytes.

Techniques: Immunohistochemistry