Figure 2.
RhoGAP19D is required for the integrity of the epithelial layer.(A) Comparison of the domain structure of Drosophila RhoGAP19D with its orthologues, human ARHGAP23/21 and C. elegans PAC-1. RhoGAP19D contains PDZ and GAP domains but lacks a PH domain. (B) Diagram showing the CRISPR-induced mutations in RhoGAP19D. The mutations generate proteins that lack the GAP domain but still contain the PDZ domain. (C) A stage 7 rhogp19d mutant egg chamber showing invasion of the follicular epithelium into the adjacent germline. The mutant cells (white arrows) invade between the nurse cells at the anterior of the egg chamber. Phalloidin staining is shown in green and DAPI in blue. (D) Graph showing the number of wild-type and rhogap19d mutant follicle cells (FC) per egg chamber at stage 8. (E) Invading rhogap19d mutant cells (marked by the loss of RFP; magenta) maintain their adherens junctions (white arrows) and epithelial organization, as shown by E-cadherin–GFP expression (DAPI; blue). (F) PH3 staining (green) of mitotic cells (white arrows) in a stage 5 egg chamber containing a rhogap19d mutant clone (marked by the loss of RFP; magenta). There is no increase in cell division in the mutant clone. (G) Quantification of the number of mitoses in wild-type and rhogap19d mutant egg chambers during stages 4 and 5. (H) mNeonGreen-RhoGAP19D (green) localizes to the lateral domain of the follicle cells and is slightly enriched at the adherens junctions (white arrows) stained with Cno (white) and DAPI (blue). RhoGAP19D protein in the germline was depleted by RNAi. (I) Surface view of wild-type cells expressing mNeonGreen-RhoGAP19D. (J and K) mNeonGreen-RhoGAP19D localizes normally in p120 catenin308/+ and p120 catenin308 homozygous cells. (L and M) mNeonGreen-RhoGAP19D recruitment to lateral domain is almost lost in cells expressing α-catenin RNAi (L) and is strongly reduced (white arrows) in shotgunIG29 mutant clones (marked by the loss of RFP; magenta; M and M'). (N and N') mNeonGreen-RhoGAP19D localization is not affected in lgl4 mutant clones (marked by the loss of RFP; magenta) compared with wild-type cells. Scale bars, 10 µm.

RhoGAP19D is required for the integrity of the epithelial layer. (A) Comparison of the domain structure of Drosophila RhoGAP19D with its orthologues, human ARHGAP23/21 and C. elegans PAC-1. RhoGAP19D contains PDZ and GAP domains but lacks a PH domain. (B) Diagram showing the CRISPR-induced mutations in RhoGAP19D. The mutations generate proteins that lack the GAP domain but still contain the PDZ domain. (C) A stage 7 rhogp19d mutant egg chamber showing invasion of the follicular epithelium into the adjacent germline. The mutant cells (white arrows) invade between the nurse cells at the anterior of the egg chamber. Phalloidin staining is shown in green and DAPI in blue. (D) Graph showing the number of wild-type and rhogap19d mutant follicle cells (FC) per egg chamber at stage 8. (E) Invading rhogap19d mutant cells (marked by the loss of RFP; magenta) maintain their adherens junctions (white arrows) and epithelial organization, as shown by E-cadherin–GFP expression (DAPI; blue). (F) PH3 staining (green) of mitotic cells (white arrows) in a stage 5 egg chamber containing a rhogap19d mutant clone (marked by the loss of RFP; magenta). There is no increase in cell division in the mutant clone. (G) Quantification of the number of mitoses in wild-type and rhogap19d mutant egg chambers during stages 4 and 5. (H) mNeonGreen-RhoGAP19D (green) localizes to the lateral domain of the follicle cells and is slightly enriched at the adherens junctions (white arrows) stained with Cno (white) and DAPI (blue). RhoGAP19D protein in the germline was depleted by RNAi. (I) Surface view of wild-type cells expressing mNeonGreen-RhoGAP19D. (J and K) mNeonGreen-RhoGAP19D localizes normally in p120 catenin308/+ and p120 catenin308 homozygous cells. (L and M) mNeonGreen-RhoGAP19D recruitment to lateral domain is almost lost in cells expressing α-catenin RNAi (L) and is strongly reduced (white arrows) in shotgunIG29 mutant clones (marked by the loss of RFP; magenta; M and M'). (N and N') mNeonGreen-RhoGAP19D localization is not affected in lgl4 mutant clones (marked by the loss of RFP; magenta) compared with wild-type cells. Scale bars, 10 µm.

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