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. 2011 Jan 21;6(1):e14553. doi: 10.1371/journal.pone.0014553

Figure 3. Knockdown of hjv does not significantly impair hepcidin expression at 55 hpf.

Figure 3

A–L. Whole mount in situ hybridization at 55 hpf for hepcidin (blue arrow) (A–C, G–I) and foxa3 (D–F, J–L), as a marker for the liver (arrowhead) and intestine (black arrow). Compared to controls (A,D), induction of bmp2b by heat shock in tg(hsp70: bmp2b) embryos (B,E) increased hepcidin expression. Treatment with dorsomorphin from 28–55 hpf in WT embryos abrogated hepcidin expression, without affecting liver size (C,F). Knockdown of hjv by a morpholino blocking translation (G,J), or by a non-overlapping morpholino targeting a splice acceptor site (H,K), did not significantly change hepcidin expression, but slightly reduced liver size. Knockdown of hjv in tg(hsp70:bmp2b) embryos failed to prevent strong hepcidin expression following induction of bmp2b (I,L). N = 10–30 embryos per group. M. The effect of hjv knockdown on bmp2b-induced hepcidin transcript levels assessed by quantitative realtime RT-PCR. Embryos were injected with hjv MO2 at the one-cell stage followed by heat shock (HS) at 48 hpf and fixation for RNA extraction at 55 hpf. N. Electrophoresis of RT-PCR products, which were designed to amplify the targeted splice site, confirmed an 80 basepair alteration in transcript size, consistent with aberrant splicing of the hjv transcript in the morphants.