Fig. 3.
Generation of Pde1c-Egfp-BAC ES cells. (A) Sagittal section of P7 Pde1c-Egfp-BAC transgenic mice shows the expression of Pde1c gene in the cerebellar granule cells, cortical Cajal Retzius cells, olfactory bulb, and pontine nucleus. ES cells were derived from F1 Pde1c-BAC 129SvJ heterozygous blastocysts. (B) Bright-field and fluorescence images showing expression of the EGFP marker of Pde1c-Egfp EBs cultured in FGF8b/RA after 4 days (16 DIV). (C) Culturing the ES cells in medium supplemented with FGF8b, WNT1, WNT3a, BMP7, and GDF7 for 4 days (8 DIV) induced expression of the granule neuron markers MEIS1 (red, D) and MATH1 (E), the cerebellar territory marker EN1 (F), and cerebellar granule neuron markers ZIC1 (G), PAX6 (H), and NEUROD (I). (Scale bar: 25 μm for B–I.)