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Supplementary Figure 4 (A) Translation-blocking antisense morpholino oligonucleotides were used to knock-down Pgam5 in Xenopus laevis embryos. Pgam5 MO1 and Pgam5 MO2 target two non-overlapping sites in pgam5.S transcripts at and close to the translation start respectively. The alignment shows the binding sites of MO1 and MO2 to the pgam5.S mRNA and base exchanges in the Pgam5 5-mismatch Control MO. (B) Alignment of Pgam5 MO1 and Pgam5 MO2 to pgam5.L. Pgam5 MO1 aligns with only two mismatches also to pgam5.L and therefore MO1 can be expected to suppress also translation from pgam5.L. The sequence in pgam5.L corresponding to the Pgam5 MO2 binding site contains seven nucleotide exchanges, therefore Pgam5 MO2 is not expected to affect pgam5.L translation. (C) Injection of either Pgam5 MO1 or Pgam5 MO2 into both blastomeres at the two-cell stage resulted in knock-down of Pgam5 at stage 11.5. (D) Representative phenotypes of embryos injected with 0.4 pmol Pgam5 MO2 or co-injected with MO-insensitive pgam5 RNA as indicated. Co-injection of pgam5 RNA rescues the phenotype of Pgam5 morphant embryos. Frequencies of phenotypes from at least three independent experiments are summarized in the graph; the total numbers of embryos are provided below each column. The χ2-test was used for statistical analysis (** p-value < 0.01). Image published in: Rauschenberger V et al. (2017) Copyright © 2017. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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