IMAGE

Fig. S1

ID
ZDB-IMAGE-131211-13
Source
Figures for Zhen et al., 2013
Image
Figure Caption

Fig. S1 The transcription and expression of d2eGFP and DsRed recapitulate that of endogenous scl-β and scl-α. (A) 5′- RACE experiment to determine the transcription start sites of DsRed and d2eGFP. The position of respective 5′-RACE primer set is indicated (arrows; R1 and R2 for DsRed; G1 and G2 for d2eGFP). Line 1 and line 2 show the 5′-RACE PCR products of DsRed and d2eGFP. M, 1 kb DNA ladder. The sequence result of 59-RACE PCR products indicates that the DsRed transcript starts from the scl-α transcription initiation site and contains the non-coding exon 1, DsRed and SV40 poly(A) sequence; the d2eGFP transcript starts form the scl-&beta initiation site and contains the non-coding sequences of exon 2, Scl-β coding sequences of exon 2, 3 and part of exon 4 (black box), followed by the d2eGFP and SV40 poly(A) sequence. The asterisks indicate the translation initiation site of DsRed and d2eGFP, respectively. The d2eGFP protein is translated as a chimeric protein fused with the N-terminal 75 amino acids of Scl-β. (B) The expression of d2eGFP and DsRed in hematopoietic stem and progenitor cells in the caudal hematopoietic tissue (CHT) of 3 dpf Tg(scl- β:d2eGFP; scl-α:DsRed) larvae. D2eGFP is observed with nucleus restriction. Scale bar: 5 μm. CV, caudal vein. (C) Upper panel shows the double immunohistochemistry staining of d2eGFP and Scl-β (detected by Ab-Scl-C) in the anterior lateral plate mesoderm, where only Scl-β is expressed. Lower panel shows the double whole-mount in situ hybridization (WISH) of DsRed and scl-α (detected by scl-5′ probe) in the ICM of 22 hpf Tg(scl-β:d2eGFP;scl-α:DsRed) embryos. Arrows indicate the colocalized cells. Embryos are shown in lateral views with anterior to the left. Scale bars: 20 μm.

Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Development