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Fig. S9
ephb4b requirement for RhoA activation and MLC2 phosphorylation at the DFCs boundary. (A) ephb4b requirement for RhoA activation at the DFCs boundary. 100 pg GST-rhotekin-RBD mRNA was injected into Tg(sox17:GFP) embryos at the one-cell stage to label active RhoA (GTP-RhoA), followed by injection with 10 ng cMO or ephb4b-MO2 injection. Embryos were collected at 75% ES for immunofluorescence staining. GST-Rhotekin-RBD (red) protein was enriched at the DFCs (green) boundary (indicated by arrows) in control embryos but not in ephb4b morphants. The ratio of embryos with the representative pattern was indicated. (B) ephb4b requirement for MLC2 phosphorylation at the DFCs boundary. Tg(sox17:GFP) embryos were injected with 10 cMO or ephb4b-MO2 at the one-cell stage and collected at 75% ES for immunofluorescence with anti-p-MLC2 (red) and anti-GFP antibodies.