Figure 6.

VAMP5 is an integral membrane protein. Total membranes derived from myotubes were extracted by different reagents as indicated. The extracts (S) and the membrane pellets (P) were then analyzed by immunoblot to detect VAMP5. As shown, VAMP5 is not extracted by PBS (lanes 1 and 2), 1 M KCl (lanes 3 and 4), 150 mM sodium bicarbonate (pH 11.5) (lanes 5 and 6), or 2.5 M urea (lanes 7 and 8) but is extracted effectively by detergents such as 1% Triton X-100 (lanes 9 and 10) and 1% DOC (lanes 11 and 12).