Fig. 3. Dgo membrane association requires fz. Panels show single confocal sections; anterior is leftwards and the equator is at the bottom (MF is at the left edge of each panel, except in A where it is on the right, marked by the red arrowhead). Mutant tissue is marked by absence of ß-gal staining or GFP (blue). White/yellow arrows indicate examples of clusters with PCP protein localization in R3/R4 cell pair (equivalent to row 3 or 4, see also Fig. 2A) and arrowheads in C-E indicate the asymmetric R4-like enrichment (around row 5-6). Green channel: anti-Dgo (in A-E). Red channel: Fz-GFP in C and D; Pk in panels E and F. Clones of null alleles of the respective genes are shown. (A) fz^– clone anterior to MF, which is marked with red arrowhead. Strikingly, also anterior to the MF, Dgo is delocalized in fz^R52 tissue [Dgo is apically localized in all wd-type cells (blue), ahead of the MF]. (B) fz^– tissue posterior to MF; apical membrane-associated Dgo localization is absent in fz^– cells (examples in wild-type area are indicated by white arrows). Dgo is enriched at membranes between fz⁺ and fz^– cells (e.g. vertical arrowhead). (C) dgo^– clone anterior to MF. There is no change in apical Fz-GFP localization in mutant tissue [compare with wild-type cells (blue)]. (D) dgo^– tissue posterior to MF. Apical membrane-associated Fz-GFP localization is not affected in dgo^– cells (examples of preclusters in wild-type and mutant areas are indicated by white arrows). (E) Pk localization in fz^– tissue posterior to MF. While apical levels of Pk localization are unaffected, the characteristic asymmetric PCP localization pattern is not observed. (F) Pk localization in dgo^–: apical localization pattern of Pk in mutant tissue is unchanged. (G,H) Dgo localization in pk^– and stbm^– clones: the Dgo pattern is as in wild type. Although Dgo displays the R4 asymmetry as in wild type, the R4-like enrichment to either equatorial or polar side is randomized [e.g. in the equatorial cell (red arrowhead), reflecting chirality flips], mimicking the adult phenotype of random chirality. (I) pk^–, stbm^– tissue: apical Dgo localization is reduced and characteristic R4-like pattern is not resolved.