Fig. 5.

Loss of Tsc1 delays the differentiation of NSCs and RGCs. a Confocal microscopy of EGFP-positive (EGFP⁺) and tdTomato-positive (tdTomato⁺) NT cells in a ROSA ^mT/mG mouse NT. Maximum intensity projections from Z-stacks is shown. b–d FACS of EGFP⁺ NT cells in b, c GFAP-Cre;ROSA ^mT/mG (control) vs. GFAP-Cre;ROSA ^mT/mG ;Tsc1 ^ff embryos or d Nestin-Cre;ROSA ^mT/mG (control) vs. Nestin-Cre;ROSA ^mT/mG ;Tsc1 ^ff embryos. b Representative FACS plots of EGFP⁺ or tdTomato⁺ NT cells. c, d GFAP-Cre;ROSA ^mT/mG (control-black) vs. GFAP-Cre;ROSA ^mT/mG ;Tsc1 ^ff embryos (blue) (c) or Nestin-Cre;ROSA ^mT/mG (control-black) vs. Nestin-Cre;ROSA ^mT/mG ;Tsc1 ^ff embryos (blue) (d). (i) Change in the percentages of EGFP⁺nestin⁺ NT cells from E15.5 to E18.5 embryos. (ii) Charts representing EGFP⁺nestin⁺ NT cells at E18.5. (iii, iv) Change in the percentages of (iii) EGFP⁺nestin⁺GFAP⁺ or (iv) EGFP⁺GFAP⁺ NT cells from E15.5 to E18.5 embryos. Data represent means ± s.e.m.. Error bars are defined as means + s.e.m. *P ≤ 0.05, ***P ≤ 0.001 t-test. Data are representative of two experiments, n_{GFAP-Cre;ROSAmT/mG} (E15.5 n = 5, E18.5 n = 3), n_{GFAP-Cre;ROSAmT/mG;Tsc1ff} (E15.5 n = 5, E18.5 n = 7) (c (i–iv)); n_{Nestin-Cre;ROSAmT/mG} (E15.5 n = 4, E18.5 n = 5), n_{Nestin-Cre;ROSAmT/mG;Tsc1ff} (E15.5 n = 7, E18.5 n = 5) (d (i, ii)); n_{Nestin-Cre;ROSAmT/mG} (E15.5 n = 4, E18.5 n = 5), n_{Nestin-Cre;ROSAmT/mG;Tsc1ff} (E15.5 n = 6, E18.5 n = 5) (d (iii, iv)) embryos