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. 2020 Feb 27;11:1105. doi: 10.1038/s41467-020-14855-3

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a, b Experimental timeline (a) and schematic diagram (b) illustrating the use of GFAP::Cre reporter mice to investigate the astrocyte-to-neuron conversion process in the striatum induced by NeuroD1 + Dlx2 (FLEx-NeuroD1-P2A-mCherry and FLEx-Dlx2-P2A-mCherry). c Typical confocal images showing the mCherry⁺ cells (NeuroD1 + Dlx2) co-stained with GFAP and NeuN at 7 dpi (left column), 28 dpi (middle column), and 56 dpi (right column). Scale bar: 20 µm. Insets show a typical cell with different markers. Scale bar: 4 µm. d Confocal images of mCherry⁺ cells (NeuroD1 + Dlx2) co-stained with S100β and NeuN at 7, 28, and 56 dpi. Scale bar: 20 µm. Inset scale bar: 4 µm. e, f Quantified data showing a gradual transition from astrocytes to neurons over the time course of 2 months in the GFAP::Cre mice after injection of NeuroD1 and Dlx2 viruses. Note that besides a decrease of astrocytes and an increase of neurons among NeuroD1 and Dlx2-infected cells, about 40% of the infected cells were caught at a transitional stage at 28 dpi, which showed both GFAP-negative and NeuN-negative (e, gray bar) or both S100β-negative and NeuN-negative (f, gray bar). Also note that the time course of astrocyte-to-neuron conversion is slower in GFAP::Cre mice compared with that induced by GFAP::Cre AAV2/5, both in combination with AAV2/5 FLEx-NeuroD1-P2A-mCherry and FLEx-Dlx2-P2A-mCherry. Data are shown as mean ± SD.