Figure 1.
Ft and Crb regulate Ex independently. (A–B′′′) Loss of Crb does not affect Ft and residual Ex remains at the apical membrane. XY (A–A′′′) and transverse (B–B′′′) confocal micrographs of third instar wing imaginal discs containing crb11A22 mutant clones (marked by absence of RFP), with Ex staining (gray in A, A′′′, B, and B′′′), Ft staining (gray in A′ and B′ and green in A′′′ and B′′′), and RFP (gray in A′′ and B′′ and red in A′′′ and B′′′). (C) A model representing the current understanding of Ft and Crb dependent regulation of Ex. Red dashes within the Ft ICD represent the conserved regions A–F. (D–E′′′) Overexpression of Ft within crb mutant tissue rescues apical Ex. XY (D, D′, E, and E′) and transverse (D′′, D′′′, E′′, and E′′′) confocal micrographs of third instar wing imaginal discs containing MARCM crb11A22 clones without UAS-expression (D–D′′′) or expressing UAS-Ft::HA (E–E′′′). Clones are marked by GFP (green in D′, D′′′, E′, and E′′′) and are stained with Ex (gray) and Ft (visualized by HA staining, red in E′ and E′′′). (F) Quantification of the ratio between apical Ex inside versus outside the MARCM clone normalized to the wild-type (wt) tissue. Data points represent an average of a single disc (n ≥ 8 per genotype) with the mean and SD indicated. **P = 0.0018 using an unpaired T test. (G–H′′′) Loss of Crb and Ft have an additive effect causing dramatic loss of apical Ex. XY (G–G′′′) and transverse (H–H′′′) confocal micrographs of third instar wing imaginal discs containing ft5-5 (marked by the absence of GFP—gray in G′ and H′, green in G′′′ and H′′′, and by green asterisks) and crb11A22 mutant clones (marked by absence of RFP—gray in G′′ and H′′, red in G′′′ and H′′′, and by red asterisks), with Ex staining (gray in G, G′′′, H, and H′′′). ft5-5 is a remake of ftfd and is a null allele. Double mutant clones are marked by absence of GFP and RFP and by yellow asterisks. (I) Quantification of the ratio between apical Ex inside indicated clone verses outside the clone. All quantification was performed on the genotype used to create double clones. Data points represent an average of a single disc (n = 6 per genotype) with the mean and SD indicated. *P = 0.0177, **P = 0.0054, and ***P < 0.0001 using one-way ANOVA with a Tukey’s post-hoc test. All XY images are orientated as dorsal up and all transverse images are apical up. Clonal boundaries are marked by yellow dotted lines. Scale bars are 10 µm.

Ft and Crb regulate Ex independently. (A–B′′′) Loss of Crb does not affect Ft and residual Ex remains at the apical membrane. XY (A–A′′′) and transverse (B–B′′′) confocal micrographs of third instar wing imaginal discs containing crb11A22 mutant clones (marked by absence of RFP), with Ex staining (gray in A, A′′′, B, and B′′′), Ft staining (gray in A′ and B′ and green in A′′′ and B′′′), and RFP (gray in A′′ and B′′ and red in A′′′ and B′′′). (C) A model representing the current understanding of Ft and Crb dependent regulation of Ex. Red dashes within the Ft ICD represent the conserved regions A–F. (D–E′′′) Overexpression of Ft within crb mutant tissue rescues apical Ex. XY (D, D′, E, and E′) and transverse (D′′, D′′′, E′′, and E′′′) confocal micrographs of third instar wing imaginal discs containing MARCM crb11A22 clones without UAS-expression (D–D′′′) or expressing UAS-Ft::HA (E–E′′′). Clones are marked by GFP (green in D′, D′′′, E′, and E′′′) and are stained with Ex (gray) and Ft (visualized by HA staining, red in E′ and E′′′). (F) Quantification of the ratio between apical Ex inside versus outside the MARCM clone normalized to the wild-type (wt) tissue. Data points represent an average of a single disc (n ≥ 8 per genotype) with the mean and SD indicated. **P = 0.0018 using an unpaired T test. (G–H′′′) Loss of Crb and Ft have an additive effect causing dramatic loss of apical Ex. XY (G–G′′′) and transverse (H–H′′′) confocal micrographs of third instar wing imaginal discs containing ft5-5 (marked by the absence of GFP—gray in G′ and H′, green in G′′′ and H′′′, and by green asterisks) and crb11A22 mutant clones (marked by absence of RFP—gray in G′′ and H′′, red in G′′′ and H′′′, and by red asterisks), with Ex staining (gray in G, G′′′, H, and H′′′). ft5-5 is a remake of ftfd and is a null allele. Double mutant clones are marked by absence of GFP and RFP and by yellow asterisks. (I) Quantification of the ratio between apical Ex inside indicated clone verses outside the clone. All quantification was performed on the genotype used to create double clones. Data points represent an average of a single disc (n = 6 per genotype) with the mean and SD indicated. *P = 0.0177, **P = 0.0054, and ***P < 0.0001 using one-way ANOVA with a Tukey’s post-hoc test. All XY images are orientated as dorsal up and all transverse images are apical up. Clonal boundaries are marked by yellow dotted lines. Scale bars are 10 µm.

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