The hematopoiesis defects induced by Ints5 knockdown are rescued by smad1 and smad5 RNA. (A-L) Whole-mount in situ hybridization to detect scl expression in 12 hpf embryos injected with control morpholinos (A,B), ints5 morpholinos (C,D), smad1 RNA (E,F), smad5 RNA (G,H), ints5 morpholinos with smad1 RNA (I,J), or ints5 morpholinos with smad5 RNA (K,L). Black arrowheads indicate the anterior limit of scl expression in the ICM; open arrowheads indicate scl expression in the RBI. A,C,E,G,I,K show dorsal views of embryos with anterior to the top; B,D,F,H,J,L show lateral views of embryos with dorsal to the right. (M) Histogram showing the percentage of injected embryos with wild-type-like (blue) or reduced (magenta) scl expression. The total number of embryos for each staining is >180, and all experiments were carried out three independent times (see Table S1 in the supplementary material). (N-S) May-Grunwald Giemsa staining to show RBC differentiation in embryos injected with control morpholinos (N), ints5 morpholinos (O), smad1 RNA (P), smad5 RNA (Q), ints5 morpholinos with smad1 RNA (R), or ints5 morpholinos with smad5 RNA (S) at 2 dpf. (T) Histogram showing the percentage of RBCs with normal (blue) or arrested (magenta) differentiation in injected embryos at 2 dpf. The total number of red blood cells counted for each sample is >200, and all experiments were carried out three independent times. Scale bars: 50 μm in A,M; 10 μm in N.
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