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C-cadherin remains at the cell surface after FLRT3
expression. A) The effect of FLRT3 expression on the accessibility
of C-cadherin to trypsin treatment of the cell surface. Blastomeres
collected from stage 9 embryos that had been injected with either
GFP RNA (as control) or FLRT3 RNA (0.2 ng) at the 4 cell stage
were either mock-treated or treated with trypsin/EDTA. Samples
were analyzed by western blotting against C-cadherin and
FLRT3. Duplicates were independent experiments. B) The effect
of activin treatment, FLRT3 expression, or PAPC expression on
the accessibility of C-cadherin to cell surface biotinylation. Stage
9 blastomeres, which were either untreated (Con), treated with 5
ng/ml activin for 1 hr, or from embryos that had been injected
with 0.2 ng of FLRT3 RNA (FLRT3) or 1.5 ng of FL-PAPC
(PAPC) at 4-cell stage, were incubated with surface biotinylation
reagent, NHS-sulfo-s-s-Biotin. The labeled surface proteins were
pulled down with neutravidin beads and analyzed by western
blotting against C-cadherin. Each lane of triplet is an independent
experiment. C) The quantification of the percentage of
biotinylated C-cadherin in samples from B. D) Anti-C-cadherin
confocal immunofluorescence microscopy of the dorsal marginal
zone cryosection of a stage 12 embryo. ec = ectoderm; me =
mesoderm; en = endoderm; yp = yolk plug.
Found at: doi:10.1371/journal.pone.0008411.s002 (9.14 MB TIF) |
