Targeted Folr1 knockdown in neural and non-neural tissues

Targeting of Folr1-MO to neural or non-neural tissues was based on cell fate maps (Moody, 1987). For microinjections, embryos were placed in 6% Ficoll in 0.1×MMR. To target the neural plate, 3.6-6 pmol Folr1-MO or CMO were injected into the dorsal animal blastomere at the eight-cell stage, or dorsal medial and dorsal lateral animal blastomeres at the 16-cell stage, 1.8-3 pmol per blastomere. To target either medial or lateral neural plate, 1.8-3 pmol of Folr1-MO was injected into the dorsal medial or dorsal lateral animal blastomere, respectively, at the 16-cell stage. To target the mesoderm, Folr1-MO was injected into the ventral medial animal, dorsal medial vegetal, ventral medial vegetal and ventral lateral vegetal blastomeres at the 16-cell stage, 3 pmol per blastomere. To target the non-neural ectoderm, the ventral lateral animal blastomere at the 16-cell stage was injected with 3 pmol Folr1-MO.