Figure 6. Lentiviral-Cyclin E transduction rescues ASPM^Δ deficiencies.

(a) Distribution of apical mitotic cleavage plane orientation of E16.5 anaphase/ telophase cells, lentiviral transduced for 48 h. χ² statistical analyses confirmed that distribution of horizontal:apicobasal:oblique ASPM^Δ divisions was significantly different than control and wasn’t rescue by lentiviral-Cyclin E transduction. Panels on right show representative axes of cleavage plane orientations. (b) Measurement of proportions of C.C exit, 24 h after BrdU pulse labeling and 48 h after in utero lentiviral transduction. (c) Cyclin E overexpression rescued the ASPM^Δ abnormal P-N cell divisions (Fig. 1j). Freshly isolated E12.5 telencephalic NPC were infected with lentiviral particles. Transgene expression was evaluated 24 h later by CFP, and only blue cells were analysed for clonal pair-fate. (d) Coronal cortical sections, lentiviral transduced at E14.5, were stained for Tuj1. Graph shows best-fit plots of the VZ fractions represented by the Tuj1⁻ region relative to the entire cortical wall radial thickness (see Fig. 1g) versus post transduction survival time. A two-way ANOVA indicates significant main effects between various genotypes and treatments (green). (e) E18.5 coronal cortical sections 96 h following lentiviral transduction stained for Cux1 and RC2. Though ASPM^Δ and WT cortices maintain comparable number of RC2⁺ cells, ASPM^Δ radial glial fibres were not extending throughout the entire cortical plate and terminated at varying positions below the Cux1 layer. (f) E18.5 coronal cortical sections 96 h following lentiviral transduction, stained for Cux1 and GFAP, demonstrating restored astrogenesis. Moreover, lentiviral-Cyclin E transduction into WT cortex accelerated astrogenesis as more radial GFAP⁺ fibers extended into the Cux1 layer (magnification of the boxed area). Bar graph demonstrates GFAP⁺ cell density within a field. All graphs display mean ± s.e.m. *P < 0.05, **P < 0.0005, ***P < 0.00001 by Student’s t-test. Scale bars, 5 µm (a), 20 µm (c), 50 µm (e,f).