ELMOD3 accumulates at actin-based structures.

<p>(A–B) GFP-ELMOD3 localizes on actin structures (elongated microvilli due to co-expression of espin) but the deafness-causing ELMOD3 mutant does not. (A) CL4 epithelial cells were co-transfected with GFP-ELMOD3 and un-tagged Espin expression vectors. Rhodamine phalloidin (red) was used to reveal F-actin and highlight the microvilli on the CL4 cell surface. GFP-ELMOD3 was efficiently targeted to the microvillar actin bundles and the cell membrane. (B) GFP-ELMOD3 harboring the p.Leu265Ser human deafness-associated mutation retained weak microvillar targeting and cell membrane localization; the protein remained in the cytoplasm and accumulated in the nucleus. (C–D) Representative results of the transfection of P2 C57BL/6J mouse inner ear sensory epithelial explants with expression vectors that encoded either GFP-ELMOD3 or p.Leu265Ser ELMOD3. The <i>x–z</i> and <i>y–z</i> plane projections are also presented (lower panels). (C) An inner hair cell (IHC) in the mouse organ of Corti that was transfected with GFP-ELMOD3 reveals localization along the length of stereocilia (arrowhead) and throughout the hair cell body. (D) Mouse organ of Corti hair cell that was transfected with GFP-ELMOD3 (p.Leu265Ser). No concentration and only negligible fluorescence is observed in the stereocilia (arrowhead), and mutated ELMOD3 remains in the cytosol. Scale bar applies to all panels and is 10 µm.