Skip to main content
. 2020 Apr 6;48(9):5065–5080. doi: 10.1093/nar/gkaa194

Figure 2.

Figure 2.

Transuterine microinjection of ASO-29 into the otic vesicle restores harmonin mRNA and protein expression. (A) ASO delivery by transuterine microinjection into the lumen of the left otic vesicle is validated by observation of the endolymphatic and cochlear ducts post-injection (compare to Figure 1A, E11.5). PHV: primary head vein. Scale bar: 1 mm. (B) ASO-29 localizes to inner hair cells (IHC, short bracket), outer hair cells (OHC, long bracket) as well as the Hensen's cell region (H, lateral to the third row of OHCs) and the pillar cell region (P, in between IHCs and the first row of OHCs) at P30. Asterisks indicate missing OHCs in the untreated mutant. Het: heterozygote. Scale bar: 20 μm. (C) Correctly spliced harmonin mRNA was detected in the P30 cochlea of ASO-29-treated heterozygous and mutant mice with a primer set that also detects the cryptically spliced form. (D) Harmonin protein expression was observed in stereociliary bundles of IHCs and OHCs of the cochlear apex in ASO-29-treated mutants at P30. Phalloidin labels filamentous actin. Arrow indicates a misshapen bundle in the ASO-29 treated mutant. Scale bar: 10 μm. (E) Higher magnification of OHCs showing harmonin expression in the bundles. Arrow indicates a misshapen bundle in the ASO-29 treated mutant. Scale bar: 5 μm. Fluorescent images are representative of n = 3 per treatment group in (B), (D) and (E). For splicing assessment, n = 4 per group in (C).