Fig. 4: S1PR1 in the SVZ-NSCs is essential for maintaining neurogenesis and ependymal planar polarity.

a Immunofluorescence images for S1PR1 (green), GFAP (cyan), SOX2 (red) and DAPI (blue) in the SVZ of control and S1pr1^ΔNestin mice (left) with mRNA (middle; n  =  5/group) and protein levels (right; n  =  4/group) of S1PR1 in NSCs from control and S1pr1^ΔNestin mice. Scale bars, 10 μm (enlarged, 5 μm). b–d Representative immunofluorescence images and quantification of SOX2⁺GFAP⁺ cells (b), MASH1⁺ cells (c), and DCX⁺ cells (d) in the SVZ (n  =  5/group). Scale bars, 30 μm (enlarged, 15 μm). e Representative immunofluorescence images of SOX2⁺GFAP⁺BrdU⁺ cells (left) and quantification of SOX2⁺GFAP⁺BrdU⁺ cells and GFAP⁺BrdU⁺Ki67⁺ cells (right) in the SVZ (n  =  5/group). Scale bars, 10 μm. f Representative images (left), number (middle), and diameter (right) of primary neurospheres (n  =  5/group). Scale bars, 300 μm. g Representative immunofluorescence images and quantification of BrdU⁺ newly formed neurons in the GCL and GL in the OB (n  =  5/group). Scale bars, 30 μm. h Quantification of CB⁺ cells, CR⁺ cells, TH⁺ cells in GL (n  =  5/group). i, j The results of olfactory habituation–dishabituation test for both urine (i) and peanut butter (j) scents (Control, n = 9; S1pr1^∆Nestin, n = 10). k The latency to locate a buried food reward (Control, n = 9; S1pr1^∆Nestin, n = 10). l Quantification of BB patch area in the walls of LV (n = 5/group). m Quantification of length/width ratios of the BB patches (n = 5/group). n Quantification of BB patch displacement (n = 5/group). o Angular distribution of BB patch angles in ependymal cells (Control, n = 296; S1pr1^∆Nestin, n = 291; five animals/group). p Quantification of CSF flow in the cerebral aqueduct (Control, n = 7; S1pr1^∆Nestin, n = 8). q Quantification of velocity of microbeads propelled by cilia at the surface (n = 4/group). r Quantification of total ventricle (left) and LV (right) volume (Control, n = 7; S1pr1^∆Nestin, n = 8). a–h, k–n, p-r Two-tailed student’s t test. i, j Two-way ANOVA for repeated measures. o Two-sided Watson U² test. All error bars indicate s.e.m. All data analysis was done at 4-month-old mice. Source data are provided as a Source Data file.