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Fig. S1. Primary cilia and Shh signaling are unaffected in hmmr morphants. Embryos at the eight cell stage (st.) were injected into one dorsal animal blastomere with hmmr morpholino (MO), asterisk indicates injected side. (A) Schematic representation of a transverse hemisection of the neural tube at the forebrain level at st. 21. (B, C) Primary cilia as detected by immunofluorescent labeling of acetylated (ac.) α-tubulin (Tuba4a; red) and γ-tubulin (Tubg1; green). DNA staining by Hoechst 33342 to detect nuclei (cyan). Rhodamine-conjugated dextran was used as lineage tracer (red background in C). Z-projection of confocal laser scanning images taken on uninjected control side (B) and hmmr MO-injected side (C) as indicated in (A). (D) Statistical analysis of cilia length; ns, not significant. Number of cilia analyzed from a total of five individuals indicated above each plot. (E, F) Transversal histological sections at the diencephalon level of embryos subjected to whole-mount in situ hybridization at st. 20. Probes detecting shh (E) and ptch1 (F) show no differences in gene expression between injected and uninjected side despite obvious morphological phenotype (compare position of arrowheads indicating the border between neural and non-neural ectoderm on injected and uninjected side). |
