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Figure 3


Fig. 3. Alternative A-site and C-site splicing affect apical and basolateral localization of PMCA2 isoforms in HCs. (A-F) Explant HCs were transfected with various EGFP-tagged PCMA2 isoform constructs and examined by immunofluorescence. An apical surface-scan of the stereocilia hair bundle (scanning plane 2 drawn in the HC schematic in G; actin labeled in red) and transverse view of the basolateral plasma membrane (plane 1 drawn in G) of a transfected HC expressing a transgene encoding for each of the EGFP-tagged PMCA2 isoforms (green) are shown in green and red, and merged channels. Arrowheads indicate strong light-scattering signal from the 1-µm-diameter gold bullets collected at 530-550 nm. Bars, 5 µm. Constructs are illustrated on the right. (G) Basolateral cross-sectional scan taken around the midpoint of the transfected HC is also shown on the right of the upper panel (scanning plane 3). (H) Summary of our data. a, a-tail; b, b-tail; AP, apical PM localization; BL, basolateral PM localization.





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