Fig. S3

A second morpholino (MO2) was designed against the e17i17 splice junction resulting in an in-frame deletion of exon 17 (corresponding to aa 666–691), leading to disruption of the CRD (A; sequence shown in Supplementary Table S2). Injection of MO2 (4–8 ng/nL) into 3.2:gfp embryos resulted in mis-routing of the epibranchial projections, similar to MO1. sVII, sIX and sX are the projections of the facial (gVII), glossopharyngeal (gIX) and vagus (gX) ganglia, respectively. Note the disruption of sX afferents and lack of hindbrain plexus. All images are oriented anterior to the left and dorsal at top. Scale bars = 100 μm. (B) Verification of effectiveness of MO2. cDNA was synthesized from four 48 hpf embryos injected with MO2 and a control, uninjected 48 hpf embryo. Primers E16.F and E18.R were designed (see Supplementary Table S2) to amplify a 280 bp fragment in control embryos (last lane on right, **). In MO2 injected embryos (lanes 1–4), a 200 bp fragment (*) was amplified, indicating a loss of exon 17 (80 bp). There is a non-specific PCR artifact at 150 bp (†), which appears in all reactions.