Both Ft and Ds suppress tissue growth by regulating localization and abundance of Dachs at the junctional cortex. (A–G) Representative wing discs stained with phalloidin showing the growth defects of the following genotypes: wild-type (A), ds^UAO71 (B), dachs^GC13 (C), ds^UAO71ft^G-rvdachs^GC13/ds^UAO71ft^fddachs^GC13 (D), ds^UAO71ft^G-rvdachs^GC13/ds^UAO71ft^fd (E), ft^G-rv/ft^fd (F), and ds^UAO71ft^G-rv/ds^UAO71ft^fd (G). Panels F and G were stitched together because of their large size (see Materials and methods). (H) Expression of ban3-GFP, a Hippo pathway reporter, compared between ft^G-rv/ft^fd (or ds^UAO71ft^G-rv/ft^fd) cells and ds^UAO71ft^Gr-v cells in the wing disc. ban3-GFP is more highly expressed in ds^UAO71ft^G-rv cells than in ft mutant cells. ds ft clones are marked by the absence of Ds staining. (I) XZ section indicated by the blue dashed line in H. Red asterisks indicate ban3-GFP expression in peripodial cells. (J) Comparison of Dachs:GFP levels and subcellular localization in wild-type, ds^UAO71, and ft^fd mutant cells. (J′) In ds mutant cells, Dachs:GFP levels were slightly increased and primarily localized to the junctional cortex (J and J′). (J″) In ft mutant cells, Dachs:GFP levels increased more than in ds cells and localized both at the junctional cortex and to non-junctional puncta (J and J″). Clone boundaries and genotypes were determined by anti-Ft staining (Fig. S1 A). (K and K′) In comparison to ft cells, ds ft mutant cells displayed even more Dachs:GFP at the junctional cortex but lacked the non-junctional puncta. (L and M) IB analysis showing Dachs levels in imaginal tissue from wild-type (number matches the n for each of the following genotypes), ds^UAO71(n = 5), ft^G-rv/ft^fd (n = 16), and ds^UAO71ft^G-rv/ds^UAO71ft^fd (n = 9). Dachs abundance increases slightly in ds but is much greater in ft and ds ft imaginal tissue. (M) Relative Dachs levels normalized to α-Tubulin. ****P < 0.0001, *** 0.0001 < P < 0.001, ns not significant (two-tailed unpaired t test between each genotype). (N) Localization of Ds and Dachs:GFP in the absence of Ft. Dachs:GFP was highly localized to the junctional cortex and colocalized with Ds in non-junctional puncta (yellow arrows). Scale bars: (A–G) 100 µm, (H–K and N) 5 µm. In this and all subsequent figures, fluorescently tagged proteins were viewed by native fluorescence. Antibody staining is indicated by an “α” symbol before the protein name. Yellow dashed lines indicate clone boundaries in this and subsequent figures (except where noted otherwise). Source data are available for this figure: SourceData F1.