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Fig. 4

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ZDB-FIG-240416-77

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Kuil et al., 2023 - ATP5PO levels regulate enteric nervous system development in zebrafish, linking Hirschsprung disease to Down Syndrome

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Fig. 4

Overexpression of ATP5PO impairs neuronal differentiation and production of ATP levels. A. Representative images of undifferentiated (T0) and differentiated (T72) control SH-SY5Y and SH-SY5Y-ATP5PO cells. Phalloidin in red, TUJ1 in green and DAPI in blue. A differentiated neuronal morphology was only observed in control cells. Moreover, TUJ1 expression was reduced in SH-SY5Y-ATP5PO cells when compared to SH-SY5Y. Scale bar represents 50 μm. B. RT-qPCR data showing increased expression of neuronal markers (TUJ1 and GAP43) in control SH-SY5Y cells 72 h after differentiation, but not in SH-SY5Y-ATP5PO cells. Mean of two technical replicates is visualized. (two-way ANOVA TUJ1: effect of ATP5PO overexpression p < 0.01, effect of differentiation p < 0.05, and interaction effect between genotype and differentiation p < 0.01; GAP43: effect of ATP5PO overexpression p < 0.0001, effect of differentiation p < 0.0001, and interaction effect between genotype and differentiation p < 0.0001) C. HuC/D stainings performed at 4 dpf showed that overexpression of ATP5PO in zebrafish, leads to reduced neuronal differentiation. D. ATP levels in control SH-SY5Y cells increase, as expected, upon induction of differentiation, whereas in SH-SY5Y-ATP5PO cells there is a decrease in ATP levels; **p < 0.01, ****p < 0.0001. A–B and D Average of three independent experiments is visualized. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

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Reprinted from Biochimica et biophysica acta. Molecular basis of disease, 1870(3), Kuil, L.E., Chauhan, R.K., de Graaf, B.M., Cheng, W.W., Kakiailatu, N.J.M., Lasabuda, R., Verhaeghe, C., Windster, J.D., Schriemer, D., Azmani, Z., Brooks, A.S., Edie, S., Reeves, R.H., Eggen, B.J.L., Shepherd, I.T., Burns, A.J., Hofstra, R.M.W., Melotte, V., Brosens, E., Alves, M.M., ATP5PO levels regulate enteric nervous system development in zebrafish, linking Hirschsprung disease to Down Syndrome, 166991, Copyright (2023) with permission from Elsevier. Full text @ BBA Molecular Basis of Disease