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. 2020 Nov 3;8(11):471. doi: 10.3390/biomedicines8110471

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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NDM29-driven differentiation of neuroblastoma cells The expression of neuron-specific protein markers in NDM29-overexpressing cells was evaluated by IF: (A) Nestin, NES; (B) Tubulin β3 Class III; TUBβ3, (C) Doublecortin, DCX and (D) Microtubule Associated Protein 2, MAP2 were detected and their relative fluorescence intensity was calculated (see Material and Methods Section), scale bars 20 µm; (E) Decrease of colony forming showing the progressive lack of malignancy from Mock to S 1.1 cells cultured in the presence of morphogens. Statistical analysis was performed excluding S1.1M cells that did not form colonies in methylcellulose; (F) The increase of population doubling-time correlates to the increase of NDM29 expression level. Statistical analysis was performed excluding S1.1M cells because of their fully mature, non-proliferating stage; (G) FACS analysis of GD2⁺ cells in different cell populations shows a linear reduction of GD2⁺ cells in clones overexpressing NDM29 ncRNA. Statistical significance is displayed as p < 0.05 (*), p < 0.01 (**).