NPF RNAi by themselves and in combination lead to different degrees of gastrulation defects. (A) Micrographs from time-lapse DIC movies in eight different backgrounds, with time indicated on the left from the MSx cell division. E lineage cells are outlined and pseudocolored in green. Gastrulation defects (Ea or Ep cell dividing before being fully covered by neighboring cells) are indicated with white arrowheads. An enclosed outline and the absence of arrowheads indicate that endodermal precursors became internalized at the 2E stage, as seen in wild-type embryos. Scale bar: 5 µm. (B) The bar graph (left) and Venn diagram (right) summarize the effects of NPF RNAi, both individually and in combination, on gastrulation. The heat map represents the different penetrance levels, with darker colors indicating a higher percentage of gastrulation defects. Fisher’s exact test was used for categorical data in B. *P < 0.05, ****P < 0.0001.