Fig. 6
Notch signaling in KA′′ specification. (A) Wild type embryos at 18 hpf were double labeled with tal2 (red) and hes5, her12, her2, or her4 (green). tal2-expressing KA′′ cells (arrows) lack expression of hes5, her12, her2, or her4. Note that hes5, her12, her2, and her4 are expressed in only a subset of LFP cells, which might reflect the oscillating nature of these genes in neural progenitor cells as previously reported [49]. Dorsal views are shown. (B) hsp-Gal4; UAS-NICD embryos and non-transgenic control embryos were heat-shocked at 10 hpf, and stained at 25 hpf for the expression of tal2 and nkx2.9 (top panel). Embryos were treated with DMSO or compound E at 13 hpf, and stained at 25 hpf for the expression of tal2 and nkx2.9 (bottom panel). tal2 and nkx2.9 staining is shown in lateral and dorsal views, respectively. Brackets indicate the extent of the LFP domain in lateral views, and arrows denote the two rows of LFP domains in dorsal views. (C) Quantification of KA′′ specification under different conditions. KA′′ cells were scored by the expression of tal2 at 25 hpf. Note that the fewer data points in compound E treatment at 13 hpf is due to the fact that most compound E-treated embryos had many tal2-positive cells in close clusters which prevents reliable scoring. Data shown therefore underestimate the total number of KA3 cells under this condition. (D) Ptc1-Kaede embryos were treated with DMSO or compound E at 14 hpf, photoconverted at 25 hpf, and stained with the GABA antibody (blue) at 37 hpf. Compound E treatment induced ectopic GABA-positive cells in the LFP domain (arrows), and abolished the expression of Ptc1-Kaedegreen (middle panel). Scale bars: 20 μm in A, D and 50 μm in B. |