Fig. 3: MT promoted the neural differentiation potential of hiPSCs by regulating MT receptor MT1/2. | Cell Death Discovery

Fig. 3: MT promoted the neural differentiation potential of hiPSCs by regulating MT receptor MT1/2.

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. 3

A Expression of MT1(12 KDa) and MT2(40 KDa) was determined by western blot after 7 days of Luzindole treatment in hiPSCs. The comparison between the target protein and the reference protein β-Actin(42KDa) revealed significant results. B The morphology of EB differentiated on days 3 and 5 after the combined treatment of Luzindole with MT. Scale bars, 50 μm. C, D The statistics of EB clone diameter and formation efficiency on day 5. E-H Expression of pluripotency marker (Nanog) and three germ layer markers (Pax6, Gata6, EOMES) was determined by qRT-PCR. I, J Expression of pluripotency marker (Nanog) and NSC markers (Pax6, Nestin) was determined by qRT-PCR. K Immunofluorescence staining of NSC markers (Pax6, Nestin). Scale bars, 50 μm. (L) The morphology of differentiated neurons with varying concentrations of Luzindole on 10 μM MT. Scale bars, 20 μm. M The statistics of number of branches per NF in spontaneously differentiated neurons. N, O Immunofluorescence staining of neuron markers (NFL, NeuN) and the differentiation efficiency statistics. Scale bars, 50 μm. P The morphology of differentiated DA neurons with varying concentrations of Luzindole on 10 μM MT. Scale bars, 20 μm. Q The statistics of number of branches per NF in directed differentiated DA neurons. R–T Immunofluorescence staining of DA neuron markers (TH, EN1, NFL, NeuN) and the differentiation efficiency statistics. Scale bars, 50 μm. U Immunofluorescence staining and quantitative analysis of DA neuronal aggregations. White arrows, neuronal aggregation. Scale bars, 10 μm.*P < 0.05, **P < 0.01, ***P < 0.001 vs. NC. #P < 0.05, ##P < 0.01 vs. MT 10 μM.

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