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Blood, Vol. 110, Issue 5, 1492-1501, September 1, 2007
SNAP-23 and syntaxin-2 localize to the extracellular surface of the platelet plasma membrane
Blood Flaumenhaft et al.
110: 1492
Supplemental materials for: Flaumenhaft et al, Vol 110, Issue 5, 1492-1501
Files in this Data Supplement:
- Figure S1. Analysis of anti-Gαq antibody labeling in intact and permeabilized platelets by flow cytometry (PDF, 10.8 KB) -
Platelets (20 µl; 0.5-1 × 108/ml) were incubated with no addition, 100 µM SFLLRN, or 0.2 µM PMA as indicated. Samples were then incubated with buffer alone (non-permeabilized) or 3 µg/ml digitonin for 30 minutes (permeabilized), stained with an IgG directed at Gαq (black) or non-immune IgG (white), and analyzed by flow cytometry. Neither antibody stained non-permeabilized platelets significantly. These results demonstrate that membrane integrity is maintained in intact platelets during analysis and confirm that the intact membrane prevents staining of intracellular proteins. Increased staining of permeabilized platelets with anti-Gαq antibody relative to non-immune antibody confirmed reactivity of the Gαq IgG. Error bars represent the standard deviation of 3 independent experiments.
- Figure S2. Exposure of intact platelets to botulinum toxin C light chain results in formation of a syntaxin-2 cleavage product (PDF, 47.9 KB) -
Intact platelets (1 × 108) were incubated in the presence or absence of 5 µg/ml botulinum toxin C light chain (BoNT C) and subsequently analyzed for syntaxin-2 by immunoblot analysis. The formation of a syntaxtin-2 cleavage product is observed following incubation with botulinum toxin C light chain.
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