FACS isolation of SVZ niche cells.

<p>(A–D) FACS plots of dissociated SVZ cells from <i>Gli1^CreER/+;hGFAP-GFP;R26^tdTomato/+</i> mice. (A) Gate was set using wild-type mice as a negative control. (B) Gate setting for GFP using <i>hGFAP-GFP</i> mice. (C) Gate setting for tdTomato using <i>Nestin-Cre;R26^tdTomato/+</i> mice. (D) FACS plot of the isolation of GFP+/tdTomato+ cells (NSCs), GFP−/tdTomato+ cells (TAPs), and GFP+/tdTomato− cells (astrocytes) from <i>Gli1^CreER/+;hGFAP-GFP;R26^tdTomato/+</i> mice. (E-F) FACS plots of dissociated SVZ cells from <i>FoxJ1-Cre;R26^YFP/+</i> mice. (E) Gate setting for a negative control using wild-type mice. (F) FACS plot of the isolation of YFP+ ependymal cells. (G-H) FACS plots of dissociated SVZ cells from <i>Tie2-GFP</i> mice. (G) Gate setting for a negative control using wild-type mice. (H) FACS plot of the isolation of GFP+ endothelial cells. (I-N) Validation of FACS-Isolated NSC niche cells by immunofluorescent staining. Nuclei were stained with Hoechst 33258 (blue). (I) GFAP, Sox2, and Nestin label NSCs (GFP+/tdTomato+). (J) Mash1 and Sox2 label TAPs (GFP−/tdTomato+). (K) GFAP and S100β label astrocytes (GFP+/tdTomato−). (L) S100β and CD24 label ependymal cells (YFP+). (M) CD31 labels endothelial cells (GFP+). (N) Quantification of immunocytochemical validation. Data represent the ratio of total antibody marker expressing cells to transgenic marker expressing cells. The results are sum of at least 2 independent experiments.