XB-IMG-158892
Xenbase Image ID: 158892
Figure 1: adam13 cleaves 792 pcdh8l. (A) Schematic representation of full length pcdh8l.
A Flag tag (DYKDDDDK) was introduced just before the first cadherin repeat (EC1).
The yellow arrow indicates the predicted cleavage site of adam13 based on the molecular
weight of N- and C- terminal fragments. Red and blue asterisk indicate N and O
glycosylation site respectively. The monoclonal antibody 2F4 was produced against the
cytoplasmic domain (Red). (B) Western blot from transfected Hek293T cells.
Glycoproteins from the conditioned media were purified on concanavalin-A agarose. The
cleavage fragment was detected using the N-terminus Flag tag using the mAb M2. Total
pcdh8l and adam13 were detected using mAb 2F4 and mAb 4A7, respectively. The Pro
(P) and Metalloprotease (M) active forms of adam13 are indicated. (C) Histogram
representing the percentage of embryos lacking fluorescent neural crest cell in the
migration pathway. (D) Photographs of representative embryos with or without
fluorescent migrating neural crest cells. Embryos were injected at the one-cell stage with
a morpholino targeting adam13 (10 ng MO13). Messenger RNA for RFP alone or
combined with the different constructs was injected at the 8-cell-stage in a dorsal animal
blastomere. Each injection was compared to RFP injected in control embryos in which
RFP positive cranial neural crest cells have successfully migrated into the branchial and
hyoid arches (0% inhibition). N=number of embryos scored from three or more
independent experiments. Error bars represent standard error to the mean. One-way
ANOVA was performed to determine statistical significance. Statistically significant at
*p < 0.05, ***p < 0.005. Image published in: Khedgikar V et al. (2017) © 2017, Khedgikar et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |