Fig. 8

Functional assessment of temporally expressed genes in border cell migration. Validation of differentially expressed genes in border cell migration using RNAi. (A-C) Pattern of c306-GAL4, driving UAS-LacZ (cyan), in stages 9–10 pre-, mid-, and post-migratory border cells and a few additional follicle cells. DAPI (gray) labels all nuclei in the egg chambers. (D-K) Knockdown of candidate genes driven by c306-Gal4; genotypes are c306-Gal4; tsGAL80 > UAS-RNAi. (D) Migration of border cells was scored based on the percentage of border cells that completed their migration (last quartile of the migration distance; 76–100%, blue) by stage 10 of oogenesis, by which time migration should be complete, or if the border cells stalled along the migration pathway, shown as the quartile distance migrated away from the anterior tip of the egg chamber (0–25%, yellow; 26–50%, pink; 51–75%, green) as shown in the schematic. (E) Quantification of migration defects for genes from the transcriptome analyses, along with the negative control (mCherry RNAi) and positive controls bazooka (Baz) and Rap1 RNAi, two genes known to regulate border cell migration. Shown are results for knockdown of 8 genes that resulted in significant migration defects at stage 10 (line); Ches-1-like (asterisk) RNAi had a strong migration defect but may partly be due to off-target effects. (F-K) Images of border cell migration at stage 10 for control (F, G) and experimental RNAi (H-K) egg chambers. Border cells (arrowheads) are labeled with E-cadherin (magenta); DAPI (gray) labels the nuclei of all cells. (F) Complete migration in a stage 10 mCherry RNAi control egg chamber. (G) RNAi knockdown of a positive control, Baz, to show representative migration defects. (H-K) Migration defects in Arf51F (H), serrano (sano; I), Bsg (J), and Cip4 (K) RNAi egg chambers. Anterior is to the left in all images. Arrowheads mark the position of border cells. All scale bars represent 10 μm. RNAi knockdown for each gene/line was performed in triplicate, with an average of 111 egg chambers per replicate. Exact N values, raw data and reagents used are available in Supplemental Data 10