Fig. 6

Non-motile OHCs are still sensitive to HPβCD. a Cytocochleograms of WT and 499-prestin-KI mice after HPβCD treatment (4000 mg/kg or 8000 mg/kg). Age and sex of mice are indicated. b-c. Immunofluorescent images of the OC from a P35 male WT (b) and a P28 male 499-prestin-KI (c) one week after a single 4000 mg/kg HPβCD injection. Images were obtained ~ 27% of the distance from apex to show the beginning of the OHC lesion boundary. Cochlear whole mount sections were stained with anti-N-mprestin antibody (green) and phalloidin-Alexa 546 (red). Scale bars, 100 μm. d Cholesterol pull-down assay (n = 4). Cell lysates containing membrane fractions from stable Sf9 cells expressing WT-prestin-V5-GFP or 499-prestin-KI-V5-GFP were incubated with unconjugated control beads or cholesterol-conjugated beads, separated on SDS-PAGE, and probed with anti-V5. Both WT-prestin-V5-GFP and 499-prestin-KI-V5-GFP are pulled-down with cholesterol-conjugated beads but not with unconjugated beads