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.