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Fig. S2 Mxtx2 induction of ndr2 transcription is independent of protein synthesis. (A-I) Embryos were injected with 50 pg gfp RNA, 25 pg eng-mxtx2 RNA or 25 pg vp16-mxtx2 RNA as indicated and divided into untreated (A-C,G-I) or cycloheximide-treated (D-F) groups. Cycloheximide treatment (50 μg/ml) was initiated at the 64- to 128-cell stage (2 to 2.25 hpf), in order to leave sufficient time for the injected RNAs to be translated into protein after RNA injection at 0.5 hpf. All embryos were fixed when control embryos reached the 30% epiboly stage (4.7 hpf) and stained for ndr2 expression. The embryos in A-F were treated in parallel; the embryos in G-I were treated in parallel to the embryos shown in Fig. 2F-H. Animal pole views are shown throughout. Fractions indicate the number of equivalent outcomes/number of embryos observed.