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Fig. S6 When Sox3 is coinjected with FGF3 or Gsc, gaps in ectopically-induced Chd expression coincide with the region of highest sox3 expression. Embryos were injected with 50 pg of sox3 plus 50 pg of either fgf3 RNA (A,B) or gsc RNA (C,D) and analysed for chd expression (blue/purple) at 4.5 hpf. Sox3 and gsc protein was detected by virtue of the HA tags they carried, using a brown peroxidase substrate. In each case the predominant region of sox3 overexpression corresponded with a gap in the region of ectopically-induced chd expression although there was often some overlap where deeper chd-expressing cells appeared to be overlaid by weaker sox3 overexpressing cells nearer the surface. Viewed from animal pole.