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. Author manuscript; available in PMC: 2015 Apr 19.
Published in final edited form as: Methods Mol Biol. 2011;741:271–283. doi: 10.1007/978-1-61779-117-8_18

Fig. 18.1.

Fig. 18.1

The endocytic recycling of CFTR. Polarized CFBE4lo- cells were grown on 24-mm Transwell permeable membrane supports for 7 days. A, Endocytosis and recycling experiments were performed as described in text and detailed in figure. B, Representative blots from endocytic and recycling assays. Cells from all four lanes were cooled to 4°C and biotinylated. The amount of biotinylated CFTR remaining in the plasma membrane after GSH treatment at 4°C (lane 2) was subtracted from the amount of CFTR remaining biotinylated after warming to 37°C and GSH treatment (lane 3) to determine the amount of endocytosed CFTR. Cells from lane 3 were then warmed at 37°C to allow recycling of endocytosed CFTR before a second GSH treatment. CFTR recycling was calculated as the difference between the amount of biotinylated CFTR after the first (lane 3) and second (lane 4) GSH treatments.