Fig. 2

iPSC differentiation toward otic neuronal progenitors (ONPs). A Representative phase-contrast photomicrographs of differentiation steps from iPSC toward ONP. Note that hi-iPSCs exhibit the same morphological changes during differentiation when compared to wt-iPSCs. Bars 100 μm. NNE: Non-neuronal ectoderm; PPE: pre-placodal ectoderm; eONP: early-stage ONP; lONP: late-stage ONP. B Representative immunocytochemistry photomicrographs of iPSC-derived lONPs, evidencing expression of the neuronal markers Nestin, β-III Tubulin (TUBB3), and neurogenic differentiation 1 (NeuroD1). The images also show expression of the otic markers PAX8, GATA-3, EYA-1, and PAX-2. Bars 20 μm. C, D Flow cytometry analysis. C ONPs derived from hi-iPSCs (hi-ONP, red) continue to lack surface expression of HLA-A, HLA-B, and HLA-C molecules following stimulation with IFN-γ when compared to wild-type cells (wt-ONP, blue). Mean fluorescent intensity values are shown in (D). Bars represent means ± standard deviation from three independent experiments. **p < 0.01