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. 2014 Aug 1;3(4):162–175. doi: 10.1089/biores.2014.0019

FIG. 4.

FIG. 4.

Expression of NEUROD1, ISLET1, BRN3A, and GATA3 in the hiPSC and hESC neurospheres at 19 and 35 DIV. (A) NeuroD1, Islet1 and Brn3a are expressed during the ganglion neuron stage of AN development and Gata3 is expressed in ANs. (B-I) Using immunostaining, an increase in the expression of ISLET1 and BRN3A was observed at 35 DIV. Scale bar=100 μm (relative to all images). (B-II, B-III) Notably, there were several ISLET1 and BRN3A immunopositive cells at 35 DIV. (C-I, C-II) There was a significant increase in the percentages of hiPSC and hESC neurospheres that had ISLET1 and BRN3A immunopositive cells. Data obtained from at least three independent experiments and expressed as means±SEM of triplicates of each sample. (D-I) The qRT-PCR data show the fold changes of NEUROD1 in the cell lines at 24 and 35 DIV relative to the control cells at 19 DIV. (D-II) The qRT-PCR results indicate the fold changes of BRN3A in the cell lines at 19 and 35 DIV relative to the undifferentiated controls. (D-III) The fold changes of GATA3 in the cell lines at 19 and 35 DIV. The qRT-PCR results were presented as fold changes relative to the endogenous control, ß-ACTIN. Each point on the graph represents mean±SD (n=3). *p<0.05; ***p<0.001.