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

Figure 4.

Figure 4

Sodiumdithionite reduces BC-associated fluorescence without getting access to SAC. Cells were labeled with 4 μM C6-NBD-GlcCer or C6-NBD-SM at 37°C for 30 min. The fluorescent lipid analogues present in the basolateral PM were then depleted by incubating the cells twice for 30 min in HBBS, supplemented with 5% BSA at 4°C. Cells were subsequently washed with PBS, rewarmed, and then incubated at 18°C in HBSS + BSA for 1 h. When the cells were subsequently treated with 30 mM sodiumdithionite in HBSS (diluted from a 1 M stock solution in 1 M Tris/HCl, pH 10) at 4°C for 7 min, C6-NBD-GlcCer (b; a, phase contrast to b) and C6-NBD-SM (d; c, phase contrast to d) were absent from BC, while leaving the fluorescence in sub-apical structures unaltered (arrows). Bar, 10 μm.