FIGURE 3.

Knockdown of Cadherin-7 with a translation-blocking morpholino antisense oligonucleotide targeting Cadherin-7 effectively reduces Cadherin-7 protein in migratory neural crest cells contributing to the trigeminal ganglion. (a) Comparison of Cadherin-7 MO (Cad7 MO) sequence to the targeting sequence in the Cadherin-7 transcript and to other Cadherin transcripts with expression patterns in premigratory and/or migratory neural crest cells, the neural tube, and in the region of the forming trigeminal ganglion. (b) Premigratory neural crest cells were electroporated at the 2–3ss with either the Cadherin-7 morpholino (Cad7 MO) to allow for depletion of Cadherin-7 protein in migratory neural crest cells, or a 5 bp mismatch Cadherin-7 control MO (Ctrl MO). Embryos were re-incubated to HH15–17 after which time the trigeminal ganglion-forming region on the electroporated side of the embryo was dissected out of the embryo and pooled for lysate preparation. Immunoblotting for Cadherin-7 and β-actin (control) was performed as in (Shah, Schiffmacher, & Taneyhill, 2017), with a representative immunoblot shown. (c) Knockdown efficiency of the Cad7 MO was assessed as previously described (Shah et al., 2017), with graph revealing results of immunoblot analysis as determined by normalizing Cadherin-7 to β-actin and calculating the reduction in this normalized ratio from that obtained for the control MO-treated lysate (arbitrarily set to 1, n = 2). The mean and standard error of the mean are shown. A 50% knockdown in Cadherin-7 protein levels is noted in the Cadherin-7 MO-treated lysate compared to the control MO-treated lysate