Fig. 3

Purification and validation of FEV-EGFP reporter system-derived SN. A FACS profiles of FEV-EGFP reporter cell (H9-Clone#7)-derived cells during differentiation from day 28 to day 42. B Representative brightfield image of the purified EGFP + cells 16 h after FACS-based purification and reseeding. C, D Immunofluorescence staining (C) and quantification (D) of EGFP and 5-HT for unsorted and sorted SN at day 42 of differentiation. E, F Immunofluorescence staining (E) and quantification (F) of EGFP and 5-HT for sorted SN at day 56 of differentiation. G Extracellular 5-HT released by unsorted or sorted cells before and after treatment with EO. H Evaluation of the sensitivity of the pharmacological responses to EO by unsorted or sorted SN. I Input voltage program (protocol) and current traces evoked by 40-ms depolarizing voltage stepped from − 40 to + 30 mV in 5-mV increments. J Current–voltage curves for voltage-gated sodium and potassium currents (n = 6). Scale bar for B: 100 μm; scale bar for C, E: 50 μm. n = 3 independent experiments. All data are presented as means ± SEM. Statistical comparisons were performed by Student’s t test. *P < 0.05; **P < 0.01; ***P < 0.001, n.s.: no significance