Figure 2. LPGDS is required for placode development(a) Increasing amounts of LPGDS-MO 10 ng (+), 100 ng (++), and 1000 ng (+++) blocks translation directed by LPGDS mRNA in an in vitro coupled transcription/translation reaction. The position of markers of known molecular weight (kDa) is indicated. (b) In situ hybridization for pan-placodal and (c) placode specific genes in control and LPGDS-MO injected embryos (frontal views, dorsal to top). Arrowheads indicate reduced expression on the injected side. (d) Quantification of the results. Four independent experiments were performed. The number of embryos analyzed (n) is indicated on the top of each bar. (e) Amino acid sequences alignment showing the conserved cysteine residue (Cyst65), center of LPGDS enzymatic activity. (f) Foxi1c and Six1 expression domains are rescued in LPGDS-MO-injected embryos by co-injection of either WT or C65A mouse LPGDS mRNA. Frontal views, dorsal to top; injected side is indicated by the lineage tracer (Red-Gal). (g) Quantification of the rescue experiment. Three independent experiments were performed. The number of embryos analyzed (n) is indicated on the top of each bar. LPGDS-MO vs. LPGDS-MO+WT or LPGDS-MO+C65A mRNA injected embryos (p<0.001, Fisher exact test.); LPGDS-MO+WT mRNA vs. LPGDS-MO+C65A mRNA injected embryos show no significant differences. Scale bars, 200 μm.
Image published in: Jaurena MB et al. (2015)
Image downloaded from an Open Access article in PubMed Central. Image reproduced on Xenbase with permission of the publisher and the copyright holder.
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