Fig. 8. Detergent insolubility. MDCK cells cotransfected with syn1A and munc-18-1 (A-B') were grown on permeable filters for 5 days and then homogenized at 4°C in buffer containing either 1% (w/v) Triton X-100 or 20 mM CHAPS. Total homogenates were subjected to centrifugation at 15,000 g for 5 minutes (L.S. = low speed) or 100,000 g for 1 hour (H.S. = high speed). The supernatants (S) and corresponding pellets (P) were analyzed by SDS-PAGE. After electrophoresis proteins in the gels were either stained with Coomassie Blue (A) or transferred to filters (B) and probed with anti-munc-18-1 (M-18-1), anti-syn1A (Syn1) and caveolin 1 (Cav-1) antibodies.
