Xenbase Image ID: 159152

Figure 2. Adam13 knockdown reduces pcdh8l expression.(A) Relative mRNA expression of pcdh8l normalized to GAPDH obtained by real-time PCR. MO13 was injected at the one cell stage (10 ng) and embryos were collected at stage 20. Polyadenylated mRNA was extracted from non-injected and MO13 injected embryos (five embryos each). (B) Representative images of whole mount in situ hybridization in which MO13 (5 ng) was injected into one of the two blastomeres and non-injected side serves as control. Image shows reduced mRNA levels of pcdh8l in MO13 injected side (star, 71% of 31 embryos). (C) Western blot on glycoproteins isolated from either control (Non Injected) or knock down (10 ng MO13) stage 20 embryos (25 embryos each). MO13 efficiently prevents the translation of adam13 (A13) as well as reduces the pcdh8l protein level. A monoclonal antibody to Xenopus Ribophorin 1 (Rpn1) was used as a loading control. Error bars represent standard error to the mean (Mean ±S.E.M). One-way ANOVA was performed to determine statistical significance. Statistically significant at *p<0.05.10.7554/eLife.26898.009Figure 2—source data 1. Source data for Figure 2.Figure 2—figure supplement 1. adam13 knockdown reduces pcdh8l expression in cranial neural crest (CNC) cells.(A) Relative mRNA expression of pcdh8l normalized to GAPDH in isolated CNC explants. CNC explants (Alfandari et al., 2003) were dissected from either control embryos or embryos injected with MO13 (10 ng at 1 cell stage) at stage 17. (B) Western blot for pcdh8l and Rpn1 from CNC glycoproteins. Proteins were extracted from non-injected and MO13 injected CNC (Alfandari et al., 2003) and glycoproteins pulled down using Con-A-agarose beads. Western blot shows that MO13 at 10 ng reduces pcdh8l expression compared to non-injected. Error bars represent standard error to the mean (Mean ±S.E.M). One-way ANOVA was performed to determine statistical significance. Statistically significant at **p<0.01.10.7554/eLife.26898.008Figure 2—figure supplement 1—source data 1. Source data for Figure 2—figure supplement 1. Larger Image Printer Friendly View

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