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. 2019 Aug 7;10:1855. doi: 10.3389/fimmu.2019.01855

Figure 1.

Figure 1

Fusion of cytotoxic granules with the plasma membrane is insensitive to Tetanus toxin. (A) Bead-stimulated human CD8+ T cells transfected with GFP or TeNT-GFP as indicated. VAMP2 and VAMP7 protein levels were determined by Western blot analysis 12–16 h after transfection. (B) Expression of VAMP2 and VAMP7 protein levels relative to GAPDH in CTLs transfected with GFP or TeNT-GFP as indicated. Graphs represent means [N = 3, ***p < 0.001 (Student's t-test)]. (C) Bead-stimulated human CTLs co-transfected with either GFP or TeNT-GFP along with granzyme B-mCherry and imaged 12 h after transfection. Representative live-cell TIRFM images of CTLs in contact with an anti-CD3 coated coverslip. Fusion events indicated with open arrowheads. (D) Mean percentage of cytotoxic granule fusion in cells transfected with either GFP (n = 50) or TeNT-GFP (n = 40), p = 0.704 (Student's t-test). (E) Mean average number of granules fused per cell over time in the TIRF plane in cells transfected with either GFP (n = 20) or TeNT-GFP (n = 15), p = 0.939 (Student's t-test). Bars show mean ± SEM. Scale bar, 5 μm.