Figure 4.
Figure 4. dmim mutants show border cell migration defects and altered endocytosis. (A) Time-lapse series of confocal micrographs of early stage 9 border cell clusters expressing UAS-GMA under 306-Gal4 in wild-type, dmim mutant, and dmim mutants expressing UAS-DMIM-Myc transgene in the border cells showing the migration of the border cell clusters over ∼3 h. White arrowheads indicate the border cell cluster and the dashed line indicates the anterior end of the oocyte (the end point of border cell migration). Bar = 50 µm. (B) Quantification of the directionality index (DI) of the border cell cluster. The DI is calculated as (A − B)/(A + B), where A is the number of forward extensions and B is the number of rearward extensions. Forward and rearward extensions are defined by which direction they point away from a vertical line on the dorsal ventral axis of the egg chamber. (C) Quantification of the migration speed of the border cell cluster, independent of the direction of migration. (D) Quantification of the average lifetime of each extension from the border cell cluster. All analyses were conducted using five separate movies for each genotype. (**, P < 0.001;***, P < 0.0001; t test). (E) Time-lapse series of confocal micrographs of individual border cells expressing UAS-GMA under 306-Gal4. Red arrowheads indicate tracking of a single cellular projection over time. Images were taken every 5 min, and the area shown is a magnification of the leading edge of a single border cell within a migrating cluster. (F) Quantification of the number of small surface projections in E. Data are represented as the mean ± SEM from 10 separate time series for each genotype. (G) Quantification of the amount of FM4-64 dye uptake over time for each indicated genotype. Data are represented as the mean ± SEM from five separate time series for each genotype. (H) Live confocal imaging of border cell clusters stained with the membrane-selective dye FM4-64. Each time series shows the gradual uptake and increase of the dye at the membrane of the border cells. Bar = 5 µm.

dmim mutants show border cell migration defects and altered endocytosis. (A) Time-lapse series of confocal micrographs of early stage 9 border cell clusters expressing UAS-GMA under 306-Gal4 in wild-type, dmim mutant, and dmim mutants expressing UAS-DMIM-Myc transgene in the border cells showing the migration of the border cell clusters over ∼3 h. White arrowheads indicate the border cell cluster and the dashed line indicates the anterior end of the oocyte (the end point of border cell migration). Bar = 50 µm. (B) Quantification of the directionality index (DI) of the border cell cluster. The DI is calculated as (A − B)/(A + B), where A is the number of forward extensions and B is the number of rearward extensions. Forward and rearward extensions are defined by which direction they point away from a vertical line on the dorsal ventral axis of the egg chamber. (C) Quantification of the migration speed of the border cell cluster, independent of the direction of migration. (D) Quantification of the average lifetime of each extension from the border cell cluster. All analyses were conducted using five separate movies for each genotype. (**, P < 0.001;***, P < 0.0001; t test). (E) Time-lapse series of confocal micrographs of individual border cells expressing UAS-GMA under 306-Gal4. Red arrowheads indicate tracking of a single cellular projection over time. Images were taken every 5 min, and the area shown is a magnification of the leading edge of a single border cell within a migrating cluster. (F) Quantification of the number of small surface projections in E. Data are represented as the mean ± SEM from 10 separate time series for each genotype. (G) Quantification of the amount of FM4-64 dye uptake over time for each indicated genotype. Data are represented as the mean ± SEM from five separate time series for each genotype. (H) Live confocal imaging of border cell clusters stained with the membrane-selective dye FM4-64. Each time series shows the gradual uptake and increase of the dye at the membrane of the border cells. Bar = 5 µm.

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