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Fig. 2. Sulf1 is required for correct DV patterning in the vertebrate neural tube. Immunostaining for Nkx2.2 (A–D), Nkx6.1 (E–H), HB9 (I–L) and Isl1 (M–P) in control (A, B, E, F, I, J, M, N) and Sulf1 knockdown (C, D, G, H, K, L, O, P) X. tropicalis embryos at stages 23 (A–P) and stage 40 (A′–P′). At stage 23, the expression of Nkx2.2 is shifted ventrally in Sulf knockdown embryos (C, D; n=12, 75%) compared with controls (A, B; n=8, 100%). At stage 40, the expression of Nkx2.2 is also shifted ventrally in Sulf knockdown embryos (C′, D′; n=18, 67%) compared with controls (A′, B′; n=11, 100%). Sulf1 knockdown only leads to a small change in Nkx6.1 expression at stage 23 (G, H; n=8, 63%), compared to controls (E, F; n=8, 100%). At stage 40 the expression of Nkx6.1 is shifted ventrally (G′, H′; n=20, 70%), compared to controls (E,′ F′; n=12, 100%). HB9 staining similarly reveals differences between the stages, at stage 23, the expression of HB9 is reduced (K, L; n=12, 75%) compared with controls (I, J; n=7, 100%). Cells positive for HB9 in Sulf1 knockdown embryos increases at stage 40 (K′, L′; n=19, 69%). At stage 23, the expression of Isl1 reduced in the MN domain of Sulf knockdown embryos (O, P; n=8, 63%) compared with controls (M, N; n=12, 100%); whereas at stage 40 Isl1 is not reliably detectable in this domain in Sulf1 knockdowns (O′, P′; n=2, 100%) or in controls (M′, N′; n=5, 100%). |
