Whole-mount in situ hybridization using tbx6, floating head (flh) and chordin probes. A–L) Animal pole views of embryos at 6 hpf with dorsal to the right. A–D) tbx6 expression is normal in uninjected (A) (n = 28/28), dramatically reduced in eve/vent/vox morphant (B) (n = 65/86), substantially decreased in eve/ved/vent morphant (C) (n = 45/68), and not significantly affected in eve/vox/ved morphant (D) (n = 71/74). E–H) flh expression of uninjected (E) (n = 31/31), eve/vent/vox morphant (F) (n = 45/47), eve/ved/vent morphant (G) (n = 38/40), and eve/vox/ved morphant (H) (n = 32/35). I–L) chordin expression of uninjected (I) (n = 24/24), eve/vent/vox morphant (J) (n = 68/68), eve/ved/vent morphant (K) (n = 67/68), and eve/vox/ved morphant (L) (n = 28/28). Arrows indicate the ventral limit of chordin expression in wild-type (I) and eve/vox/ved morphant (L). Arrowheads indicate ventrally expanded chordin expression in eve/vent/vox (J) and eve/ved/vent (K) morphant embryos. Scale bar, 200 μm.
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