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Supplemental Fig. S2. Loss of function phenotype was rescued by Mst1 or Mst2 DNA injection. (A-E) After injection of 5 ng of standard control MO (Std. MO) or Mst1/2 MO (Mst1t+2s MO) into the animal pole at the 2-cell stage, embryos were injected with 20 pg of LacZ (pENL), Mst1 (pE-Mst1), or Mst2 (pE-Mst2) DNA into the vegetal pole. Coinjection of Mst1/2 MO and Mst1, or Mst2 DNA partially rescued morphological abnormality as compared to phenotype of Mst1/2 morphant (see also Fig.2). Injected embryos were cultured until stage 38. Injected MO is indicated in the upper right of each panel. Injected DNA is indicated in the lower right. (F) RT-qPCR analysis of b-globin in embryos coinjected with MO and DNA. The coinjection of standard control MO (Std. MO) and LacZ DNA did not alter the expression level of b-globin. In Mst1/2 MO and LacZ DNA-injected embryos, expression of b-globin was decreased (see also Fig. 3). As compared to LacZ DNA, Mst1, or Mst2 DNA slightly rescued decreased b-globin expression of Mst1/2 morphant. Expression levels were normalized relative to ODC. Values represent means + SEM of three independent experiments. (G) The embryonic vascular network was formed in standard control MO and LacZ DNA-coinjected embryo. (H) Impaired vascular formation in the Mst1/2 knockdown embryo could not be rescued by injection of LacZ DNA. (D,E) Coinjection of Mst1/2 MO and Mst1, or Mst2 DNA partially rescued abnormal vascular formation as compared to phenotype of Mst1/2 MO and LacZ DNA-coinjected embryos. Sample numbers are indicated in the upper right of each panel. Developmental stage is indicated in the lower left. Injected MO and DNA are indicated in the lower right all panels. Image published in: Nejigane S et al. (2013) Copyright © 2013. Reproduced with permission of the Publisher, University of the Basque Country Press. Larger Image Printer Friendly View

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