Fig. 6. Externalized membrane vesicles are not involved in GFP secretion. CHO cells transiently transfected with the plasmid encoding GFP were starved, labeled with [³⁵S]-methionine and chased for 6 hours. At the end of the chase period, the cell lysate (C) and medium (M) were collected, centrifuged at 2,000 g for 30 minutes, and each was divided into two equal aliquots. One aliquot was immunoprecipitated with anti-GFP antibodies. The second aliquot of the medium was subjected to further centrifugation at 90,000 g for 2 hours to pellet vesicles. The supernatant (S) was collected and the pellet (P) was resuspended in 1x TXSWB buffer and immunoprecipitated with anti-GFP antibodies. The samples were separated by SDS-PAGE and analyzed by autoradiography.

Return to article