FIGURE 3.

A–F, co-localization of CDH23 and EHD4 in OK cells. OK cells were transiently co-transfected with GFP-EHD4+FLAG-CDH23 (A and B), or GFP-EHD4ΔEH+FLAG-CDH23 (D and E). After 48 h, cells were fixed and incubated with mouse anti-FLAG followed by the corresponding secondary antibody. Yellow images (right column) are superimposed from green and red images, indicating the co-localization of CDH23 and EHD4 (C) at the plasma membrane as indicated by the arrow. No co-localization was found for GFP-EHD4ΔEH and FLAG-CDH23 (F). For better visualization of the co-localization, a portion of panel C is shown in the left corner of panel C at higher magnification. G–L, no co-localization between prestin and EHD4 in OK cells. EHD4 (G–I) did not co-localize with prestin, which was used as a negative control. Bar, 23.8 μm.