Figure 4.
Par-1 delocalizes slowly from the posterior despite fast turnover. (A) Two-color time-lapse images of an egg chamber at stage 9 expressing GFP::Par-1 (cyan) and Baz::mCherry (yellow) in the germline, before (−1 min) and after ablation of PFCs (red dashed circle). Scale bar, 20 µm (B) Zoom into the posterior of the egg chamber shown in A. Top: GFP::Par-1, bottom: Baz::mCherry. Following ablation, Bazooka accumulates to the posterior, while Par-1 is still present but slowly delocalizing (arrowheads). Scale bar, 20 µm. (C) Average intensity profile of GFP::Par-1 (cyan) and Baz::mCherry (yellow) signal at the posterior of stage 9 oocytes before (top graph) and 1 h after ablation of PFCs (bottom graph). The shaded region designates the SD. The x-axis origin represents the position of the polar cells (top) or center of ablation spot (bottom). (D) Ratio between Baz::mCherry intensities at the posterior and lateral membrane of the oocyte (facing main body follicle cells), as depicted in the inset image, before (blue) and 1 h after (orange) ablation. The ratio 0 signifies complete posterior exclusion, 1 and beyond denote localization. There is a significant increase in the ratio for both stages 9 and 10A, showing accumulation of Bazooka to the posterior. (E) Ratio between GFP::Par-1 intensities at the membrane of the oocyte facing posterior ablated PFCs and intact PFCs sightly lateral, as depicted in the inset image, before (blue) and 1 h after (orange) ablation. There is no significant reduction of the ratio in stage 10A, which would indicate delocalization of Par-1. Wilcoxon signed-rank test was performed to assess significance. (F) Time series of GFP::Par-1 intensity along the oocyte membrane facing ablated PFCs (y-axis) after PFC ablation, represented as heatmap. (G) Normalized signal recovery after fluorescence photobleaching for GFP::Par-1 signal at the oocyte membrane. Thick blue line is mean, the shaded region designates the SD, the dashed line is the fitted curve obtained by fitting a single exponential function. For all panels, n is the number of experiments.

Par-1 delocalizes slowly from the posterior despite fast turnover. (A) Two-color time-lapse images of an egg chamber at stage 9 expressing GFP::Par-1 (cyan) and Baz::mCherry (yellow) in the germline, before (−1 min) and after ablation of PFCs (red dashed circle). Scale bar, 20 µm (B) Zoom into the posterior of the egg chamber shown in A. Top: GFP::Par-1, bottom: Baz::mCherry. Following ablation, Bazooka accumulates to the posterior, while Par-1 is still present but slowly delocalizing (arrowheads). Scale bar, 20 µm. (C) Average intensity profile of GFP::Par-1 (cyan) and Baz::mCherry (yellow) signal at the posterior of stage 9 oocytes before (top graph) and 1 h after ablation of PFCs (bottom graph). The shaded region designates the SD. The x-axis origin represents the position of the polar cells (top) or center of ablation spot (bottom). (D) Ratio between Baz::mCherry intensities at the posterior and lateral membrane of the oocyte (facing main body follicle cells), as depicted in the inset image, before (blue) and 1 h after (orange) ablation. The ratio 0 signifies complete posterior exclusion, 1 and beyond denote localization. There is a significant increase in the ratio for both stages 9 and 10A, showing accumulation of Bazooka to the posterior. (E) Ratio between GFP::Par-1 intensities at the membrane of the oocyte facing posterior ablated PFCs and intact PFCs sightly lateral, as depicted in the inset image, before (blue) and 1 h after (orange) ablation. There is no significant reduction of the ratio in stage 10A, which would indicate delocalization of Par-1. Wilcoxon signed-rank test was performed to assess significance. (F) Time series of GFP::Par-1 intensity along the oocyte membrane facing ablated PFCs (y-axis) after PFC ablation, represented as heatmap. (G) Normalized signal recovery after fluorescence photobleaching for GFP::Par-1 signal at the oocyte membrane. Thick blue line is mean, the shaded region designates the SD, the dashed line is the fitted curve obtained by fitting a single exponential function. For all panels, n is the number of experiments.

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