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Xenbase Image ID: 148983
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Figure 4. gmnc Is Necessary and Sufficient for MCC Differentiation in Xenopus(A) End-point PCRs on stage-30 control and gmnc morphants showed mis-splicing of the Xenopus gmnc pre-mRNA.(B) Western-blot analysis using an antibody against human GMNC showed that endogenous Gmnc was depleted in the gmnc morphant embryos at stage 35 and that the injected human GMNC RNA translated to a large amount of the GMNC protein. Actin was used as a loading control.(C and D) The development of gmnc morphant embryos was delayed (observed in 93% of the 43 animals tested). Despite this, in situ hybridization revealed no overt defects in the mucus-secreting goblet cell differentiation (81%; n = 21) (C) or in the differentiation of scattered cells, which are involved in ionic homeostasis regulation (78%; n = 18) (D).(E and F) In contrast to the lack of differentiation defects in goblet cells and scattered cells, the differentiation of MCCs, the third main cell type present on the Xenopus embryonic skin, and marked by ccdc19 (encoding a ciliary protein), was significantly reduced in the gmnc morphants (87%; n = 52) (F) compared to control embryos (E).(G and H) Immuno-staining of acetylated-tubulin revealed that multiple cilia were markedly reduced in the gmnc morphants (94%; n = 34) (H) compared to control embryos (G).(I–K) High-magnification images of the surface ectoderm at stage 30 in control (I), gmnc morphants (J), or emrbyos co-injected with gmnc MO and 80 pg of human GMNC RNA (K) showed that overexpression of the hGMNC RNA not only rescued the MCC defects in the gmnc morphants but was sufficient to produce ectopic MCCs (79%; n = 14).See also Figure S4. Image published in: Zhou F et al. (2015) Copyright © 2015. Image reproduced with permission of the Publisher, Elsevier B. V.
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