Hh signaling is upregulated in Emilin3 morphant embryos. (A) Wild-type (WT) and Tg12x_Gli embryos were treated with 100 μM cyclopamine (cyc) and probed at 24 hpf by in situ hybridization for ptc1 or mCherry. (B) qRT-PCR for ptc1 and mCherry transcripts in Emilin3 morphant and control embryos (*P<0.05; n=30). (C) In situ hybridization for nkx2.2a, tbx20 and myoD1 in 24 hpf injected embryos. All images are lateral views, except for bottom panels (dorsal views). (D) In situ hybridization for shh, ehh and ntl in 24 hpf injected embryos. (E) Immunofluorescence for engrailed in 24 hpf injected embryos. Scale bars: 150 μm. (F) Quantification of engrailed-positive cells at 24 hpf (*P<0.05; n=10). Data are mean+s.e.m. Ctrl MO, control morpholino; e3a MO, emilin3a morpholino; e3b MO, emilin3b morpholino; e3a+e3b MO, emilin3a + emilin3b morpholinos; MPs, muscle pioneers; MFFs, medial fast fibers.