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. 1998 Aug 10;142(3):683–696. doi: 10.1083/jcb.142.3.683

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Semi-quantitative analysis of BCP labeling: accumulation of C₆-NBD-lipid analogues in SAC. Cells were labeled and treated as depicted in Fig. 1. A represents the distribution of NBD lipid in BC and SAC after step 1 (30 min at 37°C) while the distribution indicated in B was obtained after a subsequent back exchange and chase from BC at 18°C for 60 min (Fig. 1, step 2 and 3). In both cases, the percentage C₆-NBD-GlcCer- and C₆-NBD-SM-labeled BCP was ∼85%. Labeling of BCP was distinguished in either NBD-positive BC only, a cellular BCP fraction in which both BC and SAC were labeled (BC + SAC), or NBD-positive SAC. Data are expressed as percentage of the total number of C₆-NBD-GlcCer– (white bars) and C₆-NBD-SM–positive (hatched bars) BCP (minimum of 50/coverslip; mean ± SEM) of at least four independent experiments carried out in duplicate.

Semi-quantitative analysis of BCP labeling: accumulation of C₆-NBD-lipid analogues in SAC. Cells were labeled and treated as depicted in Fig. 1. A represents the distribution of NBD lipid in BC and SAC after step 1 (30 min at 37°C) while the distribution indicated in B was obtained after a subsequent back exchange and chase from BC at 18°C for 60 min (Fig. 1, step 2 and 3). In both cases, the percentage C₆-NBD-GlcCer- and C₆-NBD-SM-labeled BCP was ∼85%. Labeling of BCP was distinguished in either NBD-positive BC only, a cellular BCP fraction in which both BC and SAC were labeled (BC + SAC), or NBD-positive SAC. Data are expressed as percentage of the total number of C₆-NBD-GlcCer– (white bars) and C₆-NBD-SM–positive (hatched bars) BCP (minimum of 50/coverslip; mean ± SEM) of at least four independent experiments carried out in duplicate.