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Fig. 5. Effects of amotl2 expression knockdown on cell movements and
structures. (A) Migratory behavior of clonal cells. One cell of a
64-cell embryo was injected with lissamine-labeled control MO1 (cMO1) or
fluorescein-labeled amotl2MO1 (MO1). According to location of labeled
descendants at the shield stage (SS), embryos were separated into three
groups: dorsal, lateral and ventral. The sorted embryos were observed again
during early segmentation (ES) and at 24 hpf. Cells targeted by cMO1
contributed to corresponding embryonic tissues at 24 hpf (upper panel), while
MO1-targeted cells mainly clustered on the yolk at 24 hpf regardless of their
early locations (lower panel). The bar graphs show percentage of embryos with
labeled cells in embryonic tissues (wild type) or on the yolk based on four
independent experiments. The total number of observed embryos is indicated in
parentheses. (B) Effect of amotl2 expression knockdown on cell
shape. One cell of a 64-cell embryo was injected with 10 pg mRNA coding for
membrane GFP alone or in combination with either 5 ng cMO1 or MO1. The labeled
cells were observed at the end of gastrulation by confocal microscopy.
Compared to the wild-type or cMO1-injected cells, MO1-injected cells had a
round shape and lost membrane protrusions such as filopodia. (C) Effect
of amotl2 expression knockdown on F-actin. Embryos were injected with
corresponding morpholinos at the one-cell stage and subsequently stained at
the shield stage with phalloidin. Outer layer cells of flat-mounted embryos
were photographed. Notice that, in MO1-injected cells, F-actin is less
abundant in the peripheral region and not well ordered intracellularly.
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