Extended Data Fig. 2: Fip200 KO and Fip200-4A knock-in mutant rescued defective phenotypes in Tsc1^GFAPcKO SVZ.

a,b, Immunofluorescence of DCX (a), GFAP (b) and DAPI in SVZ of Tsc1^GFAPcKO and 2cKO mice at P21. Four independent experiments gave similar results. c–f, Mean ± s.e. of the percentage of β-III tubulin⁺ cells (c), NeuN⁺ cells (d), GFAP⁺ cells (e) and MBP⁺ cells (f) from differentiated neurospheres of Ctrl, Tsc1^GFAPcKO, 2cKO and Fip200^GFAPcKO mice. n = 4 (c,d), 5 (e,f) independent experiments. g, Frequency of SEN-like structure in Ctrl, Tsc1^GFAPcKO, 2cKO, 2cKI, and Fip200^GFAPcKO mice at P7, P14 and P28. The animal numbers used at different postnatal stages are indicated. (h–l) Immunofluorescence of pS6RP (h), nestin and Ki67 (i), GFAP and Sox2 (j), NG2 (k), DCX and GFAP (l) in SEN-like lesion in Tsc1^GFAPcKO brain at P21. Three independent experiments gave similar results. m, Immunofluorescence of pS6RP and DAPI in SVZ of Tsc1^GFAPcKO and 2cKI mice at P21. Five independent experiments gave similar results. n, Mean ± s.e. of the number of pS6RP⁺cells in SVZ of Ctrl, Tsc1^GFAPcKO, 2cKI and Fip200^GFAP cKI mice at P21. n = 5 independent experiments. o, H&E staining of SVZ from Ctrl, 2cKI and Fip200^GFAP cKI mice at P28. Five independent experiments gave similar results. p, Mean ± s.e. of SVZ cell number of Ctrl, Tsc1^GFAPcKO, 2cKI and Fip200^GFAPcKI mice at P21. n = 5 independent experiments. q,r, Mean ± s.e. of the number of GFAP⁺Nestin⁺ NSCs (q) and GFAP⁺Sox2⁺ NSC (r) in SVZ of Ctrl, Tsc1^GFAPcKO, 2cKI and Fip200^GFAPcKI mice at P21. n = 5 independent experiments. The dotted lines indicated the boundaries of SVZ or SEN-like lesion with LV. E, ependymal layer; LV, lateral ventricle; SEN, subependymal nodule-like lesion; ST, striatum; SVZ, subventricular zone. Scale bar, 100 μm (a,b,m,o), 40 μm (h–l). Data were analysed by one-way ANOVA with Tukey’s post-hoc test (c–f,n,p–r) or Chi-square test (g).