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. 2021 May 10;12:2594. doi: 10.1038/s41467-021-22813-w

Fig. 1. Quiescence NSCs express high levels of the Cdt1 red Fucci reporter in vivo.

Fig. 1

a Immunohistochemistry for GFAP (green) in the SVZ. mCherry Fucci reporter (red) and nuclear counterstaining with DAPI (blue). Right, detail of GFAP positive cells expressing different levels of Fucci red reporter and quantification (n = 3 independent mice). b Immunohistochemistry for GFAP (yellow), CD9 (green), mCHERRY Fucci reporter (red), and nuclear counterstaining DAPI (blue). Right, detail of GFAP positive cells with different levels of CD9 and mCherry Fucci reporter. Nuclear counterstaining with DAPI (blue). c Live imaging of the Fucci NSCs treated with EGF/FGF, BMP and BMP/FGF showing mCHERRY (red) and VENUS (green) reporters. d Flow cytometry Fucci quantification of the percentage of cells positive for Cdt1-mCherry (red) and VENUS (green), negative or low levels of fluorescence (pale pink) and double positives for both reporters (yellow) (n = 3). e qPCR for different markers (Egfr, Hes5, Mmc2, Sox2, Gfap, Id1) in the different conditions (data relative to EGF/FGF) (n = 3). f Immunocytochemistry for CD9 (yellow) in Fucci NSC line, mCherry (red) and nuclear counterstaining with DAPI (blue) (n = 3 independent NSC lines). g Quantification of the CD9 mean intensity in NSCs treated for EGF/FGF2, BMP4, and BMP/FGF2 for 3 days. Quantification by Fiji (n = 3 independent experiments per condition, average intensity of min 500 cells per group). h Number of colonies in the sorted population of Fucci cell cycle reporter based high levels of CD9 and mCHERRY levels (h-high, l-low) (n = 3). Data are shown as mean ± SEM of the indicated number of the experiments (n) (*p < 0.05; **p < 0.01; ***p < 0.001). St:striatum, LV: Lateral Ventricle. Scale bar in (a, b) is 20 μm, (c, f) is 50 μm. Scale in details:10 μm. Source data are provided as a Source Data File.