Figure 6

Knockdown of Ngn2 expression does not induce a change in laminar fate. E15.5 embryonic cortices were electroporated with plasmids encoding the YFP variant Venus and either control shRNA or shRNA targeting Ngn2. Following electroporation, organotypic slice cultures were prepared and cultured for 5 DIV. To identify any potential change in laminar fate, slices were stained for CTIP2, a molecular marker for deep layers (layers 5 to 6) and Cux1, a molecular marker for superficial layers (layers 2 to 4). (A,D,G,J) Electroporated cells were identified by immunostaining for EGFP. (B,C,E,F) CTIP2 immunostaining (B,E) and the merged images (C,F) demonstrated that very few electroporated neurons express the deep layer marker. (M) Quantification of these data shows no significant difference in the number of neurons that were immunostained for GFP and CTIP2. (H-L) Cux1 immunostaining (H,K) and merged images (I,L) show that most of the electroporated neurons express the superficial layer marker. (N) Quantification demonstrates that there is no significant difference in the number of neurons that were immunostained for GFP and Cux1. Interestingly, the location of the neurons within the cortex has no effect on the expression of laminar markers (arrowheads). NS, not significant. Error bars indicate standard deviation.