Fig. S5

scl-α MO is specific to the defects of HSC maintenance in the AGM of scl-α morphants. (A-C) The loss of HSCs is partially rescued in the scl-α morphant receiving scl-α PAC expression. (A) The scl-α PAC used for rescue. DsRed and an SV40 polyadenylation signal were inserted in exon 4 to interrupt the transcription of scl-β. To introduce normal expression of scl-α, the DNA sequences of scl exon 2, 3, 4 and an SV40 polyadenylation signal were inserted behind exon 1. (B) CD41:eGFP+ HSCs in the AGM region of live 2 dpf control embryos, scl-α morphants and scl-α morphants injected with scl-α PAC. White arrows indicate CD41:eGFP+ HSCs in the AGM. Blue arrowheads identify pronephric duct cells. Scale bar: 20 μm. (C) Statistical analysis showing the number of CD41:eGFP+ HSCs (per five somites) in the AGM of 2 dpf control embryos, scl-α morphants and scl-α morphants injected with scl-α PAC. (D-I) Double immunostaining of Cmyb-eGFP and TUNEL in the AGM of 36 hpf scl-α morphants (D-F) and control embryos (G-I). White arrows indicate three Cmyb:eGFP+ cells undergoing apoptosis in the AGM. Scale bar: 20 μm.