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Fig. 5. Quantification of hair cell differentiation and rows in wild-type and
COUP-TFI-/- cochlea after DAPT treatment. (A)
Real-time PCR analysis of myosin VIIa transcripts shows hair cell
differentiation is dependent on DAPT dose. There is a significant difference
in the COUP-TFI-/- compared with wild type at the
concentration of 5 µM (**P<0.05). (B) Western
blot confirmation of myosin VIIa protein expression after treatment with 5
µM DAPT shows an increase with DAPT treatment. The myosin band of
Kaleidoscope prestained standard is used as control. (C) Hair cell row
counts (myosin VIIa-positive inner and outer hair cells) along the length of
the cochlear duct after culture with 5 µM DAPT. Both wild-type and
COUP-TFI-/- cochleae showed significant increases in hair
cell rows after DAPT treatment (*P<0.05), and
COUP-TFI-/- mutants had a significantly greater number
than wild type in the apex (**P<0.05). (D)
Immunostained sections of DAPT-treated cochleae at the apical region show an
increase in hair cell rows in the COUP-TFI-/- mutants
(c,d) compared with wild type (a,b; note, image in D part d is an extreme case
of extra hair cells). Sections show hair cells expressing myosin VIIa (red)
and nuclei counterstained with DAPI (blue). Scale bar: 20 µm.
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