VAMP-7 Antibody

Western Blot: VAMP-7 Antibody [NBP2-32232] -

Western Blot: VAMP-7 Antibody [NBP2-32232] - VAMP7 forms a SNARE complex with SNAP-23 & Syntaxin11. (A) Model illustrating the formation of SNARE complex between VAMP7 (v-SNARE), SNAP-23, & Syntaxin11 (t- SNARE) during cytotoxic granule fusion in human CD8+ T cells. (B) Different Twin-Strep-tag fusion constructs used for pulldown assay. Twin-Strep-tag was fused at the C terminus of VAMP7 & at the N terminus of Syntaxin11 with a (GGS)x3 linker. (C) Western blot of bead stimulated human CD8+ T cells transfected with Twin-Strep-tag-tagged VAMP7, immuno-precipitated with anti-FLAG antibody & detected with antibodies against FLAG, SNAP-23, & Syntaxin11. (D) Western blot of bead stimulated human CD8+ T cells transfected with Twin-Strep-tag-tagged Syntaxin11, immuno-precipitated with anti-Syntaxin antibody & detected with antibodies against Strep-tag, SNAP-23, VAMP7, & VAMP2. As control, cells were transfected with Twin-Strep-tag construct. Ten percentage of the lysates were loaded as input. (E,F) Densitometric quantification of the Western blots shown in (C,D), respectively. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31447853), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: VAMP-7 Antibody [NBP2-32232] -

Western Blot: VAMP-7 Antibody [NBP2-32232] - Knockdown of VAMP7 strongly reduces fusion of cytotoxic granules at the IS. (A) Lysates from bead stimulated human CD8+ T cells transfected with either control or VAMP7 siRNAs (1 or 2, respectively) & blotted for VAMP7 (top) & GAPDH (bottom) as loading control. (B) Quantification of VAMP7 protein expression (in % normalized to control siRNA-treated CTLs) performed by densitometry. Bars indicate SEMs. [VAMP7-siRNA1, N = 3; ***p < 0.001 & VAMP7-siRNA2, N = 3; ***p < 0.001 (t-test)]. (C) Human CD8+ T cells co-transfected with granzyme B-mCherry along with either ns-siRNA or VAMP7-siRNA1 or VAMP7-siRNA2 & imaged 12 h after transfection. Selected live-cell TIRF microscopy images of granzyme B-mCherry in a transfected CTL in contact with an anti-CD3 coated coverslip. Fusion events are indicated with open circles (three frames shown per granule fused). (D) Mean percentage of cytotoxic granule fusion in cells transfected with either ns-siRNA (n = 66 & n = 59, respectively) or VAMP7-siRNA1 [n = 91; **p < 0.01 (t-test)] or VAMP7-siRNA2 [n = 72; ***p < 0.001 (t-test)]. (E) Mean average number of granules fused over time in the TIRF plane per cell p = 0.206 (t-test) for VAMP7-siRNA1 & **p < 0.01 (t-test) for VAMP7-siRNA2. Bars indicate mean ± SEM. Scale bar, 5 μm. (F) Calcein-based killing assay for CTLs transfected with either ns-siRNA, VAMP7-siRNA1, or VAMP7-siRNA2. Experiments were carried out in duplicate [N = 4; ***p < 0.001 (t-test)]. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31447853), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: VAMP-7 Antibody [NBP2-32232] -

Western Blot: VAMP-7 Antibody [NBP2-32232] - 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 & VAMP7 protein levels were determined by Western blot analysis 12–16 h after transfection. (B) Expression of VAMP2 & 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 & 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. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31447853), licensed under a CC-BY license. Not internally tested by Novus Biologicals.