Figure 3.
Bazooka localizes to the posterior following ablation of PFCs. (A) Two-color time-lapse images of an egg chamber expressing Baz::GFP in the germline (yellow) and the microtubule reporter Jupiter::mCherry (magenta) before (−1 min) and after ablation of PFCs. Scale bar, 20 μm. (B) Zoom-in of the posterior end of the egg chamber shown in panel A. Top: merged channels, bottom: Baz::GFP. Before ablation, Bazooka is excluded from the posterior, a region highlighted by the arrowhead in the first image of the bottom panel. After 90 min, accumulation of Bazooka is visible at the oocyte membrane facing the ablated cells (arrowheads in the last image). Scale bar, 20 μm. (C) Average intensity profile of Baz::GFP signal at the posterior of the oocyte before (blue) and after (orange) ablation. The shaded region designates the SD. The x-axis origin represents the position of the ablated region. Vertical lines denote the average width of the ablated region. Note that the increase of the signal is visible only inside the ablated region. (D) Ratio between average Baz::GFP intensities along the oocyte membrane facing ablated PFCs and lateral main body follicle cells, as depicted in the inset image, before (blue) and after (orange) ablation for three types of ablation. PFCs: ablation of PFCs, including ablation of polar cells (as in A). Nonpolar PFCs: ablation of PFCs that do not include polar cells (see Fig. S2 A). Ooplasm: ablation inside the ooplasm (see Fig. S2 D). A ratio equal to 1 means complete loss of Bazooka exclusion. Wilcoxon signed-rank test was performed to assess significance. (E) Intensity of Baz::GFP signal at the oocyte membrane facing the ablated follicle cells as a function of time, grouped in experiments where polar cells were ablated (orange, n = 5) or left intact (blue, n = 5). The thick lines represent the average of each group, thin colored lines are individual oocytes. Intensity of the ooplasm was subtracted from the Baz::GFP intensity, so that zero intensity (on average) means exclusion of Bazooka. Inset: Average curves in the first 15 min. (F) Time series of Baz::GFP intensity along the oocyte membrane facing ablated PFCs (y-axis) after PFC ablation, represented as heatmap. The ablated follicle cells included the polar cells, and the ablated region is flanked by still intact PFCs. (G) As in F, but for ablations that excluded polar cells, so that the ablated region was flanked by intact polar cells on one side (bottom of y-axis) and main body follicle cells on the other side (top of y-axis). Note the signal inflow from the top of the graph. For all panels t = 0 is the first frame after ablation, n is the number of experiments.

Bazooka localizes to the posterior following ablation of PFCs. (A) Two-color time-lapse images of an egg chamber expressing Baz::GFP in the germline (yellow) and the microtubule reporter Jupiter::mCherry (magenta) before (−1 min) and after ablation of PFCs. Scale bar, 20 μm. (B) Zoom-in of the posterior end of the egg chamber shown in panel A. Top: merged channels, bottom: Baz::GFP. Before ablation, Bazooka is excluded from the posterior, a region highlighted by the arrowhead in the first image of the bottom panel. After 90 min, accumulation of Bazooka is visible at the oocyte membrane facing the ablated cells (arrowheads in the last image). Scale bar, 20 μm. (C) Average intensity profile of Baz::GFP signal at the posterior of the oocyte before (blue) and after (orange) ablation. The shaded region designates the SD. The x-axis origin represents the position of the ablated region. Vertical lines denote the average width of the ablated region. Note that the increase of the signal is visible only inside the ablated region. (D) Ratio between average Baz::GFP intensities along the oocyte membrane facing ablated PFCs and lateral main body follicle cells, as depicted in the inset image, before (blue) and after (orange) ablation for three types of ablation. PFCs: ablation of PFCs, including ablation of polar cells (as in A). Nonpolar PFCs: ablation of PFCs that do not include polar cells (see Fig. S2 A). Ooplasm: ablation inside the ooplasm (see Fig. S2 D). A ratio equal to 1 means complete loss of Bazooka exclusion. Wilcoxon signed-rank test was performed to assess significance. (E) Intensity of Baz::GFP signal at the oocyte membrane facing the ablated follicle cells as a function of time, grouped in experiments where polar cells were ablated (orange, n = 5) or left intact (blue, n = 5). The thick lines represent the average of each group, thin colored lines are individual oocytes. Intensity of the ooplasm was subtracted from the Baz::GFP intensity, so that zero intensity (on average) means exclusion of Bazooka. Inset: Average curves in the first 15 min. (F) Time series of Baz::GFP intensity along the oocyte membrane facing ablated PFCs (y-axis) after PFC ablation, represented as heatmap. The ablated follicle cells included the polar cells, and the ablated region is flanked by still intact PFCs. (G) As in F, but for ablations that excluded polar cells, so that the ablated region was flanked by intact polar cells on one side (bottom of y-axis) and main body follicle cells on the other side (top of y-axis). Note the signal inflow from the top of the graph. For all panels t = 0 is the first frame after ablation, n is the number of experiments.

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