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Figure 4
Knockdown of discs large 5 (dlg5) in Xenopus embryos causes cell polarity and Hedgehog signalling dysfunction. (A) Representative images of dnah9 in situ hybridisation in control MO and dlg5 MO sides of a stage 28 embryo depict fainter expression signal in the dlg5 depleted side. (B) Representative transverse sections of control MO and dlg5 MO sides of a stage 28 embryo reveal the more basal location of cells expressing the dnah9 marker of MCC cell fate. (C) Representative immunofluorescence images and quantitation of AcTub-positive ciliated cells/area and Lectin-positive goblet cells/area in control MO and dlg5 MO sides of a stage 28 embryo demonstrate the decreased density of MCCs and concomitant increased density of goblet cells in dlg5 depleted Xenopus epidermis. (D) Representative WISH for foxf1 Hedgehog target expression in control MO and dlg5 MO stage 45 pronephroi demonstrate the decreased Hedgehog signalling activity in dlg5 depleted pronephroi. (E) Representative expression and quantitation of ptch1 in control MO and dlg5 MO stage 45 brain ventricles as well as representative expression and quantitation of Pax6 in control MO and dlg5 MO stage 45 fourth ventricle demonstrate a decrease in ptch1 expression and expansion of Pax6. Black dotted line delineates bounds of the brain and white dotted line delineates bounds of fourth ventricle floor. A, anterior; D, dorsal; P, posterior; V, ventral. Statistical tests carried out as two-tailed t-tests for cell density (C) and Pax6 expression (E) and unpaired t-tests for foxf1 (D) and ptch1 (E) expression with ***p<0.005 and ****p<0.0001. Bars indicate mean and SD of individual values (E—Pax6 expression) or replicate means (D and E—foxf1 or ptch1 expression). |
