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Fig. 3. | BMC Biology

Fig. 3.

From: Notch signaling is a critical initiator of roof plate formation as revealed by the use of RNA profiling of the dorsal neural tube

Fig. 3.

Activation of Notch signaling at the ventral roof plate border. al Transverse sections through the dorsal neural tube at the level of the forelimb in wildtype (WT) and Wnt1-Cre; Mib1fl/fl mouse embryos, immunostained for N1ICD (white or red) and DAPI (blue). N1ICD staining is shown in combination with DAPI (a, c, e, g, i, k) or as a single color image for clarity (b, d, f, h, j, l). Low levels of N1ICD are evident in the neural tube at E9.75 (a–d); however, at E10.5 (e–h) and E11.5 (il), N1ICD is broadly expressed throughout the dorsal neural tube in wildtype embryos, whereas this is diminished in Wnt1-Cre; Mib1fl/fl embryos. m–x Transverse sections through the dorsal neural tube at the level of the forelimb in wildtype (WT) and Wnt1-Cre; Mib1fl/fl mouse embryos, immunostained for DLL1 (red or white) and DAPI (blue). DLL1 staining is shown in combination with DAPI (m, o, q, s, u, w) or as a single color image for clarity (n, p, r, t, v, x). At all ages examined, in wildtypes, DLL1 is expressed throughout the neural tube except for its dorsal-most region corresponding to the RP (boundary demonstrated by dashed lines in n, r, v). In Wnt1-Cre; Mib1fl/fl embryos, DLL1 protein accumulates at the cell membrane due to lack of Mib1-induced endocytosis, and by E11.5 there are no DLL1-negative cells present in the dorsal-most neural tube. y, z Quantification of DLL1-negative cells in the dorsal-most neural tube at E10.5 and E11.5. At E10.5, there is no difference between genotypes, whereas at E11.5 there is a significant reduction in the number of DLL1-negative cells, and hence of the presumptive RP, in Wnt1-Cre; Mib1fl/fl embryos. N = 6; ns, not significant; ****p < 0.0001. aa Double immunostaining for Dll1 (green) and N1ICD (red) at E11.5; note that the dorsal limit of expression of both markers coincides (dotted lines). bb, cc, dd Gdf7 ISH (blue or green) and N1ICD immunostaining (red) of adjacent sections showing a side-by-side expression of both markers with no apparent overlap (dotted lines). Bar = 50 μm

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