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Fig. 2 vegfc loss-of-function leads to altered definitive hematopoiesis throughout development. (A) In vivo imaging of cd41:gfp (green; HSPCs) and flk:mCherry (magenta; vessels) depicting decreased HSPC numbers in the CHT of vegfc loss-of-function embryos at 72 hpf. (B) Quantification of cd41:gfp+ cells within the CHT, as shown in A; Cas9 control versus vegfc crispants, P=0.0023 (unpaired two-tailed t-test). (C) vegfc crispants show decreased area of rag1 expression in the thymus at 5 dpf. Black arrow highlights area of rag1 staining in vegfc crispant. (D) Quantification of the area of rag1 expression, P<0.0001 (unpaired two-tailed t-test). (E) vegfc loss-of-function increases the number of cmyb-expressing cells in the CHT compared with Cas9 control embryos. (F) Quantification of the number of cmyb-expressing cells in the CHT at 72 hpf, P<0.0001 (unpaired two-tailed t-test). (G) vegfcum18−/− mutant embryos show an increased number of cmyb-expressing cells in the CHT at 72 hpf. (H) Number of cmyb-expressing cells in the CHT at 72 hpf, P<0.0001 (unpaired two-tailed t-test). Error bars show mean±s.e.m. Scale bar: 40 µm (A).