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Fig. 4. NHE3-EGFP did not recover after photobleaching in either the apical plasma membrane or JN areas after DSS cross-linking. OK/NHE3-EGFP cells had fluorescence recovery studied after the cells were exposed to the membrane-penetrable cross-linking reagent DSS (disuccimidyl suberate) (1 mM for 1 hour at 37°C). The NHE3-EGFP fluorescence did not recover in either the apical domain or in the intracellular juxtanuclear (JN) region after photobleaching. (A) Time-course montage after photobleaching (the numbers at the bottom are seconds after the bleach) of apical membrane NHE3-EGFP in OK cells. Magnification 100x objective. (B) Quantitative fluorescence recovery of NHE3-EGFP in the apical plasma membrane. Similar results were obtained of NHE3-EGFP in the intracellular JN domain. Note, the apical membrane studied (A and B) included NHE3-EGFP over the JN plus the non-JN domain. Mobile fractions for NHE3-EGFP in Cyto-JN and the apical plasma membrane (PM) are shown in C. n=5 for Cyto-JN and n=6 for PM from two separate experiments.





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