Fig. 6.

Live-imaging analysis of Magoh-depleted interneuron progenitors reveals autonomous requirements for mitotic progression, survival and cell fate. (A) Schematic of 24 h clonal live-imaging analysis with fixed analysis of progeny cell fate (example on the right). Dashed lines in images represent cell borders. (B-G) Analyses of Nkx2.1-Cre; Rosa^{loxSTOPloxtdTomato} (control), Nkx2.1-Cre; Magoh^lox/+; Rosa^{loxSTOPloxtdTomato} (cHet) and Nkx2.1-Cre; Magoh^lox/lox; Rosa^{loxSTOPloxtdTomato} (cKO). (B) Quantification of average mitosis duration (minutes) of tdTomato⁺ cells. (C) Histogram of mitosis duration (minutes). A threshold of delayed mitosis at 50 min (dashed line) is based upon control progenitors completing mitosis within this period. (D) Quantification of the proportion of viable (gray) and non-viable (red) divisions. Non-viable divisions characterized as mitotic slippage were due to unsuccessful cytokinesis and subsequent cell death. (E,F) Quantification of the proportion of progenitor progeny surviving (gray) or undergoing apoptosis (red) amongst all progeny (E) or relative to mitosis duration (F). (G) Quantification of the proportion of progeny fate undergoing neurogenic (white) or proliferative (gray) divisions. Neuronal fate was determined post-imaging by Tuj1⁺ staining and progenitor fate was determined by Ki67⁺Tuj1⁻ staining. ANOVA with Tukey post-hoc: *P<0.05, **P<0.01, ***P<0.001 (B). χ² analysis with post-hoc Bonferroni-adjusted P-values represented by asterisks: *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 (B,D-G). ns, not significant. Error bars represent s.d. Experiments represent two live-imaging sessions, two litters; control n=3 embryos (182 cells), cHet n=3 embryos (219 cells), cKO n=2 embryos (57 cells). Scale bar: 10 µm.