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Fig. 4. DAPT treatment of cochleae in culture induces hair cell
differentiation. (A) Real-time PCR analysis of myosin VIIa
transcripts is presented in terms of average 
Ct (see Materials and
methods). A greater Ct value represents a lower transcript expression level.
Histogram and bars represent average Ct±s.d.
(*P<0.05). High DAPT doses increase myosin VIIa
expression, with significant induction at 5 µM DAPT. (B) Western
blot confirmation of myosin VIIa protein expression changes with DAPT dose.
The myosin band of Kaleidoscope prestained standard is used as control.
(C) Expression of COUP-TFI and COUP-TFII transcripts
in cochleae treated with DAPT. COUP-TFI and COUP-TFII
expression was not changed with DAPT, even at the highest concentration of 5
µM. Because DAPT can induce hair cell differentiation (in terms of myosin
VIIa expression as shown in Fig.
4A), the value is presented here after further normalization with
the myosin VIIa values. (D) Expression of Hes5 in cochleae
after culture with DAPT. Hes5 expression is reduced with increasing
concentrations of DAPT, with statistically significant differences at 2.5 and
5 µM (*P<0.05). A statistically significant
reduction was also found between COUP-TFI-/- and wild type
at 5 µM (**P<0.05).
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