Xenbase Image ID: 153358

Fig. S2. Shh MO efficiently and specifically blocks Shh function. (A) Schematic of Shh transcript with exons (blue), introns (Vs), and UTRs (gray) shown. “Shh spliceMO” (red) was designed to overlap the first splice donor site. RT-PCR primers (black) were designed flanking the splice junction to assay for correct splicing. (B) RT-PCR of embryos injected with Shh spliceMO shows dose-dependent reduction of Shh splicing. EF1α serves as a loading control. (C–J) ISH for Hh pathway target genes Patched1/2. Shh mRNA upregulates Ptc (D), while Shh MO reduces Ptc (E). Reduction by Shh MO is specific, as it can be rescued by co-injection of Shh mRNA (F). In a separate round of experiments, cyclopamine treatment shows a similar amount of Ptc reduction (H) to Shh MO (I). Combination of these reagents does not enhance the phenotype (J), indicating that maximal possible knockdown of Shh function is achieved by either Shh MO or cyclopamine alone. Image published in: Peyrot SM et al. (2011) Copyright © 2011. Image reproduced with permission of the Publisher, Elsevier B. V. Gene Synonyms Species Stage(s) Tissue ptch1.S patched, patched1, Ptc-1, Ptc1, X-ptc1, XPtc1 X. laevis Throughout NF stage 15 notochord neural plate neural tube Image source: Published Experiment + Assay Source Phenotypes and Disease Xla Wt + shh + NF15 (in situ hybridization) fig.S2.d Expression Phenotype increased amount ptch1.S expression in neural plate mislocalised ptch1.S expression in neural plate Xla Wt + shh MO + NF16 (in situ hybridization) fig.S2.e, i Expression Phenotype decreased amount ptch1.S expression in neural tube Xla Wt + Cyclopamine + NF16 (in situ hybridization) fig.S2.h Expression Phenotype decreased amount ptch1.S expression in neural tube Larger Image Printer Friendly View

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