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Fig. S1 The hsp70:cyp26a1 line efficiently inhibits RA signaling. (A) Heat shock of hsp70:cyp26a1 embryos for 1 hour starting at 6 hpf results in close proximity of the first somite to the otic vesicle and a kink at the head-trunk boundary at 24 hpf, representative of complete loss of RA signaling. (B) Whole-mount in situ hybridization for egr2a, a marker of hindbrain rhombomeres 3 and 5, illustrates absence of rhombomere 5 in hsp70:cyp26a1 embryos, caused by posterior expansion of more anterior rhombomeres. hs, heat shock; o, otic vesicle; s, first somite; 3, rhombomere 3; 5, rhombomere 5. Scale bars: 100 μm.