Expression of PAX2 in the hiPSC and hESC neurospheres at 19 and 35 DIV. (A) Pax2 is expressed in otic placodal cells during the early stages of AN development. (B) Using immunostaining, an increase in the expression of PAX2 was observed at 35 DIV. Scale bar=100 μm (relative to all images). (C-I) There was a significant increase in the percentages of hiPSC and hESC neurospheres that had PAX2 immunopositive cells. Data obtained from at least three independent experiments and expressed as means±SEM of triplicates of each sample. (C-II) The qRT-PCR data show the fold changes of PAX2 in the cell lines at 19 and 35 DIV. The results were presented as fold changes relative to the endogenous control, ß-ACTIN, and the undifferentiated stem cells. Each point on the graph represents mean±SD (n=3). *p<0.05; **p<0.01; ***p<0.001. DIV, days in vitro; DAPI, 4′,6-diamidino-2-phenylindole; SD, standard deviation; SEM, standard error of the mean.