Fig 1: Defective actin clearance and centrosome polarization at the immunological synapse in Spag6KO mice.(A,B) The immunological synapse between WT or spag6KO CD8+ CTLs (red) and P815 target cells was stained for (A) F-actin (green, white arrows, upper panel) or (B) ?-tubulin (green dots, white arrows, lower panel). (C) Quantification of actin clearance (left) and centrosome (?-tubulin) polarization and docking distance from the synapse in WT and Spag6KO CD8+ cells shown. P values are shown.
Fig 2: Reduced CTL function in Spag6KO mice.(A) Allogeneic cytotoxicity assay with decreasing ratios of WT (red) or Spag6KO (blue) CD8+ T cells to P815 target cells; (B) Soluble IFN? production by activated WT or Spag6KO CD8+ T cells (C) WT or Spag6KO (KO) CD8+ T cell proliferation in media alone (white) or with CD3/CD28 stimulation (stripes). (D) Histogram showing intracellular IFN? labeling in WT and Spag6KO CD8 T cells compared to isotype control. (E) The geometric mean of intracellular IFN? fluorescence intensity (GMFI) is significantly higher in the Spag6KO CD8s compared to the WT (F) Intracellular IFN? distribution in activated WT (upper panel) or Spag6KO (lower panel) CD8+ T cells. N = 3 – 7 per group; **p < 0.005.
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