Fig. 3

Identification of three genes in a pilot pRNAi screen of the DEG candidates. A Images showing the development of live gfp pRNAi, g26874 pRNAi, g7720 pRNAi, and g4238 pRNAi embryos. These images are related to Movie S2 (Additional file 4). Stages of development and time (h) after egg laying (AEL) are shown. Asterisks indicate the embryonic pole, closed blastopore, and posterior pole. Arrowheads indicate the rim of the forming/formed germ disc, or corresponding sites in morphologically affected embryos. The arrow indicates CM cell migration, which did not occur in the g26874 pRNAi, g7720 pRNAi, and g4238 RNAi embryos. B Effect of g26874, g7720, or g4238 pRNAi on the expression of the corresponding target gene at stage 3 and/or late stage 5, as revealed by WISH. gfp pRNAi or wild-type (WT) embryos were stained as controls. At least three embryos were stained for each treatment type. Counterstains with DAPI were displayed for the comparison between g26874 and gfp pRNAi embryos. (C) Graphs showing developmental profiling of the transcript levels (RPKM; reads per kilobase of transcript, per million mapped reads) for g26874, g7720, and g4238 in wild-type embryos, based on public datasets [36]. Two biological replicates are individually shown. The sampling times were 9 (stage 1), 15 (stage 2), 20/21 (stage 3), 25/26 (stage 4), 31/32 (early stage 5), 36/37 (late stage 5), 42/43 (stage 6), 50 (stage 7), 60 (stage 8), and 78 (stage 10) h AEL. D Results of blast searches against D. melanogaster and mouse RefSeq protein databases using predicted amino acid sequences of g26874, g7720, and g4238 as queries. The top-hit protein from each search is shown. Scale bars, 100 µm in A and B