Fig. 5. Identification of specific brain regions in brain organoids. (A) Forebrain markers (PAX6, FOXG1) and hindbrain markers (PAX2, ISL1) were quantified by RT-PCR in spheroids at 10 and 16 days of differentiation. The expression values were normalized to the β-actin expression level. Three independent experiments had been performed. Data are shown as mean ± SD. *p < 0.05, **p < 0.01. (B) Immunohistochemical staining of PAX6 and PAX2 markers in cryosections of spheroids at 16 days and 33 days. (C) Immunohistochemical staining of PAX6 and ISL1 markers in cryosections of brain organoids at 33 days. Scale bars = 50 μm.