Fig. S9. Ectopic otic vesicle formation by activation of Wnt signaling is reduced when sp8 expression is down-regulated. Ectopic otic vesicle formation was evaluated by oc90 expression. Embryos incubated with 6-bromoindirubin-3′-oxime (BIO) from stage 13/14 to stage 28 were subject to in situ hybridization analysis. Whereas control embryos showed normal oc90 expression (A), BIO-treated embryos showed increased or ectopic oc90 expression (B–D). To address whether Wnt-induced ectopic otic vesicle formation requires sp8, embryos receiving sp8 ATGMO or sp8 TALEN injection at the two-cell stage were incubated with (E) or without (F) BIO from stage 13/14 to stage 28. Interestingly, BIO-induced ectopic otic vesicles were not observed in sp8 MO or TALEN injected embryos (E), indicating sp8 is required for the Wnt-mediated ectopic otic vesicle formation. Black arrows indicate ectopic oc90 expression. In addition, we observed that oc90 expression was retained on the dkk1-injected side (H) in comparision to the uninjected side (G), suggesting that the absence of sp8 expression in dkk1- injected embryos is not due to the loss of otic vesicle tissue.
Image published in: Chung HA et al. (2014)
Copyright © 2014. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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