Fig 2: Isothiocyanate-conjugated phalloidin labels both cell membrane and ciliary bundles for comparisons of the morphologies of the WT, untreated, and low- and high-dose-treated Kcne1-/- mice at P30 (n = 4 in each group).a The morphologies of marginal cells (MCs), ciliary bundles of cochlear, and vestibular hair cells (HCs) in WT mice as labeled. b The morphologies of MCs, ciliary bundles of cochlear, and vestibular HCs in untreated Kcne1-/- mice. c The morphologies of MCs, ciliary bundles of cochlear, and vestibular HCs in low-dosage-treated Kcne1-/- mice. The ciliary bundles of vestibular HCs in the utricle and crista ampullaris (CA) were significantly preserved. d The morphologies of MCs, ciliary bundles of cochlear, and vestibular HCs in high-dosage-treated Kcne1-/- mice. The shape of the MCs and the ciliary bundles of the HCs in the cochlear and vestibular organs seemed to have recovered to normal. Red: phalloidin.

Fig 3: Comparison of Kcne1 expression levels in marginal cells (MCs) and vestibular dark cells in WT mice, untreated Kcne1-/- mice, and low- and high-dose-treated Kcne1-/- mice at P30 (n = 4 in each group).a Kcne1 expression in MCs and vestibular dark cells in WT mice. b Absence of Kcne1 expression levels in MCs and vestibular dark cells in untreated Kcne1-/- mice. c Kcne1 expression levels were found in some MCs and vestibular dark cells in low-dose-treated Kcne1-/- mice. d Extensive Kcne1 expression levels in MCs and vestibular dark cells in high-dose-treated Kcne1-/- mice. Scale bar is 50 µm and applies to all panels. Details of data quantifications are given in the text. Red: phalloidin; Green: KCNE1.

Fig 4: (A) Representative immunocytochemistries of H9c2 cells cultured in presence of glucose 11.1 mM Normal Glucosio), 33 mM glucose (High glu) and labelled with KCNE1 antibody. Representative microscopic fields for each treatment are shown with percentage of KCNE1-positive cells is represented in the graph. (B) The results are expressed as mean ± S.E.M. of the percentages of positive cell / total cell counted in each analyzed field for each treatment. **P < 0.01 Normal glu; °P < 0.05 versus High glu; °°P < 0.01 vs High glu. 40× magnification.

Fig 5: Representative merged images of MitoTracker labeling of mitochondria (red), immunofluorescence with KCNE1 antibody (green) and counterstaining of cell nuclei with Hoechst 33258 (blue) in H9c2 cells cultured in normal glucose (Normal glu) or 33 mM glucose (High glu) and exposed to DMSO 0.05 µM and BF-5m 0.01–0.025–0.05 µMMerged images did not demonstrate a significant colocalization of KCNE1 with the mitochondria in H9c2 exposed to normal or high glucose medium and receiving DMSO 0.1 µM or BF-5m 0.01–0.025–0.05 µM pretreatment. 40× magnification.