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Fig. 2. Xevl knockdown disrupts otic vesicle development. (A) Western blot analysis using a Xevl polyclonal antibody shows a marked reduction in Xevl protein production in embryos injected with XevlMO compared with embryos injected with control MO (coMO). Levels of -tubulin are unaffected by injection of the XevlMO. Numbers on right indicate molecular mass markers (kDa). (B) Quantitative analysis of Xevl knockdown and rescue experiments indicating the percentage of embryos displaying perturbed otic vesicle development. (C-G) Head region of Xenopus embryos at stage 35 (C) Diagram showing the olfactory placode (ol), lens placode (lens), otic vesicle (ov), epibranchial placodes (epi), and lateral line placodes (unlabeled). (D) XEya1 expression on the uninjected side of the embryo. (E) XEya1 expression on the XevlMO-injected side of the embryo. Otic vesicle size is reduced in 86% of Xevl-depleted embryos (n=121) with 72% exhibiting a strong reduction in size and 14% displaying a mild reduction. Otic vesicle size was unaffected in 14% of injected embryos. (F) XEya1 expression on the uninjected side of a rescued embryo. (G) XEya1 expression on the side of the embryo injected with Xevl-GFP mRNA and XevlMO. Expression of Xevl-GFP results in rescue of XEya1 expression with 43% displaying normal otic vesicles, 21% exhibiting a mild phenotype and 36% exhibiting a strong phenotype (n=77). Arrow marks the otic vesicle in D-G. |
