(A) Immunofluorescence confocal microscopic imaging from mouse ventricular myocytes, demonstrating the expression of prestin (Slc26a5, middle panel, cyan). Cells were counterstained with phalloidin-fluorescein isothiocyanate (FITC; top panel, green). The merged image is shown on the bottom panel.
(B) A negative control using ventricular myocytes isolated from Slc26a5−/−. Cells were counterstained with anti-α-actinin2 antibody (green).
(C) A cryosection from the mouse organ of Corti as a positive control, demonstrating the presence of prestin (cyan) in the lateral membrane of the outer hair cells (OHCs) but not inner hair cells (IHCs) as expected. DAPI (4’,6-diamidino-2-phenylindole) was used to stain the nuclei (blue).
(D) Immunofluorescence super-resolution STED imaging from Prestin-YFP KI mouse ventricular myocytes showing the expression of prestin using anti-GFP antibody (green). Cells were counterstained using anti-α-actinin2 antibody (red).
(E) A negative control using anti-GFP antibodies in the WT control. Cells were counterstained using anti-α-actinin2 antibody (red).
(F) A cryosection from the mouse organ of Corti as a positive control from Prestin-YFP KI mice showing the presence of prestin (green) in the lateral membrane of OHCs but not IHCs. DAPI was used to stain the nuclei. All scale bars are 10 μm.