Fig. 4. Mutation of Y520Q and Y667Q results in targeting of prestin to the plasma membrane of HEK cells, and presence of NLC in Y667Q, Y520F and Y667F.

The upper panels show HEK cells transfected with wild type prestin-YFP, and the two constructs Y520Q prestin YFP, and Y667Q prestin YFP that were fixed 48 hours after transfection. Cells were stained with antibodies to Na/K ATPase and visualized by confocal microscopy. Wild type prestin YFP, Y520Q prestin YFP and Y667Q prestin YFP all target the plasma membrane as evidenced by its co-localization with plasma membrane Na/K ATPase. The lower panel shows NLC traces of different mutations at Y520 and Y667. The actual values of NLC parameters along with cell numbers are given in supplementary material Table S1. The scale bar is 10 microns.