XB-IMG-158899
Xenbase Image ID: 158899
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Figure 5: Adam13 induces pcdh8l via AP2. (A, C, D) Relative expression of AP2
and pcdh8l in naïve ectoderm (animal cap) by real-time PCR. One-cell stage embryos
were injected with 1 ng of the various adam13 constructs and morpholinos and embryos
were collected at stage 9 to dissect animal cap cells (AC). AC were grown in 0.5X MBS
until sibling embryos reached stage 20 to 22. Messenger RNA was extracted from 10 AC
for gene expression analysis. Expression was normalized using GAPDH and compared to
the expression in non-injected AC (NI). (A) Real-time PCR data show that adam13 can
induce AP2α by more than 4 fold. Both proteolytic activity and the presence of the
cytoplasmic domain are essential for full activation. (B) Luciferase activity of AP2α
promoter in Hek293T cells show induction by adam13 and ADAM19 but not ADAM9.
For each transfection the Luciferase values were normalized to the Renilla values driven
by the CMV promoter. In these assays, the absence of proteolytic activity (A13E/A)
reduced AP2 induction only slightly, while the deletion of the cytoplasmic domain
(Cyto) prevented the activity. (C) Induction of AP2by adam13 was prevented by the
KD of AP2 (10 ng of MOAP2) but not -catenin (20 ng of Mo-catenin). (D)
Expression of AP2 in response of adam13 also depends on AP2 but not -catenin.
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,
***p < 0.001. 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 |