Fig. 1: MT regulates the differentiation potential of hiPSCs into neuroectoderm and NSCs in vitro. | Cell Death Discovery

Fig. 1: MT regulates the differentiation potential of hiPSCs into neuroectoderm and NSCs in vitro.

From: Melatonin orchestrates mitochondrial fusion dynamics-mediated WNT/β-catenin signaling to promote dopaminergic neuronal differentiation of human iPS and nerve regeneration in a MPTP-induced mouse model of Parkinson’s disease

Fig. 1

A Alkaline phosphatase staining and relative AP activity statistics. AP staining intensity was measured using the image J software.Scale bars, 20 μm. B Expression of pluripotency markers (Nanog and OCT4) in hiPSCs at mRNA level. C Relative OD values measured by CCK8 experiments reflecting the self-renewal ability of hiPSCs treated with different concentrations of MT. D An optimized protocol of EB random differentiation. E The morphology of EB differentiated on days 3 and 5. Scale bars, 50 μm. F, G The statistics of EB clone diameter and formation efficiency on day 5. H–K Expression of pluripotency marker (Nanog) and three germ layer markers (Pax6, Gata6, EOMES) was determined by qRT-PCR. L An enhanced small molecule induction scheme to differentiate hiPSCs into NSCs. M The morphology of differentiated NSC with standard neural rosettes with varying concentrations of MT on day 7. White arrows, neural rosette. Scale bars, 50 μm. N, O Expression of pluripotency marker (Nanog) and NSC markers (Pax6, Nestin) was determined by qRT-PCR. (P–R) Immunofluorescence staining of NSC markers (Pax6, Nestin, SOX1, SOX2) and the midbrain neural marker (Nurr1). Scale bars, 50 μm. S The statistics of the relative fluorescence intensity in differentiated NSCs on day 7, and fluorescence intensity data were analyzed using ZEN microscope imaging software. (T) Flow cytometry analysis of NSC markers (Pax6, Nestin, SOX2, SOX1) and the midbrain neural marker (Nurr1). U Relative OD values measured by CCK8 experiments reflecting the self-renewal ability of NSCs treated with different concentrations of MT. *P < 0.05, **P < 0.01, ***P < 0.001 vs. NC.

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