Figure 2.
Polyploidy is essential for border cell migration. (A) Egg chambers from c306-Gal4>UAS-GFP ovary. The C306-Gal4 is specifically expressed in the stalk cells (arrows) and the border cells (arrowheads). Nuclei are labeled with DAPI (blue). (B) Stage 10a egg chambers expressing CycE-RNAi in border cells cluster (upper panel) or E2f1PIP3A (a stable and active form of E2f1) in the border cell cluster (lower panel). F-actin is stained with phalloidin (red) and nuclei with DAPI (blue). (C) Quantification of border cell ploidy in the stage 10 egg chambers. Nuclear area was measured from z-stack confocal images using the central cross section (the largest image) of each nucleus. The left two quantitative data in the graph show the results of oocyte follicle cells (OFCs) and border cells from stage 10 egg chambers expressing only GFP in border cells. In stage 10 egg chambers, all follicle cells should have completed three rounds of endoreplication, corresponding to ∼16C ploidy. Compared with these controls, border cells expressing CycE-RNAi or E2f1PIP3A show nuclear areas less than half of control, indicating they lack one or two rounds of endoreplication compared with normal 16C ploidy cells. **** represents P < 0.001 (unpaired two-tailed Student’s t test); n = 30 cells from six egg chambers from c306-Gal4>UAS-GFP (GFP OFCs and GFP), c306-Gal4>UAS-CycE-RNAi, and c306-Gal4>UAS-E2f1PIP3A ovaries. (D) Quantification of border cell migration in the egg chambers from c306-Gal4 (n = 47), c306-Gal4>UAS-CycE-RNAi (n = 70), and c306-Gal4>UAS-E2f1PIP3A (n = 107) ovaries. (E) A schematic diagram of the border cell migration index. Scale bars, 50 μm. Refer to the image caption for details. Panel A: Fluorescent microscopy images showing C 306-G a l 4 greater than U A S-G F P egg chambers with D A P I labeled nuclei and marked border cells. Panel B: Fluorescent microscopy images showing C y c E-R N A i and E 2 f 1 P I P 3 A expressing border cells stained with phalloidin and D A P I. Panel C: Scatter plot showing nuclear size quantification of border cells under different conditions with statistical significance indicated. Panel D: Vertical bar graph showing migration index distribution of border cells across W T, C y c E-R N A i, and E 2 f 1 P I P 3 A conditions. Panel E: Schematic diagram showing anterior to posterior border cell migration path with migration index scale representation.

Polyploidy is essential for border cell migration. (A) Egg chambers from c306-Gal4>UAS-GFP ovary. The C306-Gal4 is specifically expressed in the stalk cells (arrows) and the border cells (arrowheads). Nuclei are labeled with DAPI (blue). (B) Stage 10a egg chambers expressing CycE-RNAi in border cells cluster (upper panel) or E2f1PIP3A (a stable and active form of E2f1) in the border cell cluster (lower panel). F-actin is stained with phalloidin (red) and nuclei with DAPI (blue). (C) Quantification of border cell ploidy in the stage 10 egg chambers. Nuclear area was measured from z-stack confocal images using the central cross section (the largest image) of each nucleus. The left two quantitative data in the graph show the results of oocyte follicle cells (OFCs) and border cells from stage 10 egg chambers expressing only GFP in border cells. In stage 10 egg chambers, all follicle cells should have completed three rounds of endoreplication, corresponding to ∼16C ploidy. Compared with these controls, border cells expressing CycE-RNAi or E2f1PIP3A show nuclear areas less than half of control, indicating they lack one or two rounds of endoreplication compared with normal 16C ploidy cells. **** represents P < 0.001 (unpaired two-tailed Student’s t test); n = 30 cells from six egg chambers from c306-Gal4>UAS-GFP (GFP OFCs and GFP), c306-Gal4>UAS-CycE-RNAi, and c306-Gal4>UAS-E2f1PIP3A ovaries. (D) Quantification of border cell migration in the egg chambers from c306-Gal4 (n = 47), c306-Gal4>UAS-CycE-RNAi (n = 70), and c306-Gal4>UAS-E2f1PIP3A (n = 107) ovaries. (E) A schematic diagram of the border cell migration index. Scale bars, 50 μm.

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