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Fig. 10. The TM2-TM3 region is sufficient for NHE7 interaction. (A) CHO cells were transiently co-transfected with HA-tagged NHE7 and GFP-tagged SCAMP2 TM2-TM3 (GFP-TM2-3) or GFP control. Cell lysates (Lys) were immunoprecipitated with anti-HA antibody (IP) and bound GFP fusion proteins were analyzed on western blot. (B) Purified GST fusion protein of NHE7 C-terminus or GST was incubated with radiolabelled GFP-TM2-3 and bound protein was detected using a PhosphorImager. (C) CHO cells were simultaneously transfected with GFP-TM2-3 or GFP alone and NHE7HA and their intracellular localization was visualized by green and red fluorescence, respectively. (D) Homogenate isolated after transient transfection with GFP-TM2-3, GFP, SCAMP2_myc or NHE7_HA was analyzed by sucrose equilibrium density centrifugation. Bar, 3 µm.

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