View larger version (49K):
[in a new window]
|
Fig. 1. Difference gel electrophoresis (DIGE) experimental scheme. Collections of
ventralized embryos at different developmental stages were compared with
lateralized embryos. (A) The general scheme for the DIGE experiments was to
prepare whole embryo extracts from ventralized or lateralized embryos. The
proteins within each extract were covalently labeled with either propyl-Cy3 or
methyl-Cy5. The labeled protein extracts were then combined and
co-electrophoresed on a 2DE gel. After electrophoresis, the gel was fixed in
methanol and acetic acid and then imaged in the fluorescent gel imaging device
at the Cy3 and Cy5 excitation wavelengths. Common protein spots have equal
amounts of red-colored Cy3 and blue-colored Cy5, which is shown here as
purple. Proteins that are unique to the Cy3-labeled sample are shown as red
spots; unique Cy5-labeled proteins are shown as blue spots. (B) Ventralized
and lateralized embryos were compared at three developmental stages:
pre-cellularization (VPC and LPC), early gastrulation
(VEG and LEG) and late gastrulation (VLG and
LLG). Shown here are three frames from time-lapse recordings of a
ventralized and of a lateralized embryo at the indicated stage. Anterior is to
the left. The double-headed arrows indicate the various comparisons that were
made. The cephalic furrow is indicated by arrowheads in the LEG and
LLG images, the cephalic furrow does not form in ventralized
embryos.
|
