Supporting Information

Transcription

1 Supporting Information Willecke et al /pnas A A' clones A'' A''' ' GFP clones ' fj- clones D D' Diap1 fj- clones Fig. S1. oundaries of and activity induce Hippo target genes. Third instar imaginal discs containing clones of cells either overexpressing or mutant for fj. Differences in or levels at clone borders induced expression of lacz enhancer trap insertions into the Hippo target genes ex (A and D) and Diap1 () or the DIAP1 protein (). ells overexpressing (A and ) are positively marked by coexpression of GFP. lones mutant for fj d1 ( and D) are marked by the absence of GFP. A show eye imaginal discs and D shows a wing disc. -Gal antibody stainings are shown in red in A D and in gray in A D. (A ) The -Gal antibody staining is shown in gray and a red line outlines the clone border. (A ) GFP is shown in gray. Anterior is to the left in all discs and arrowheads point to clone borders. Willecke et al. 1of5

2 A' clones ' clones, ft - ' clones, d - Fig. S2. boundaries require Fat and Dachs for their action. Imaginal discs with clones of cells overexpressing (marked by GFP expression in green). (A) expression induced the expression of diap1-lacz along clone borders in an eye imaginal disc. expressing clones did not induce ex-lacz in a fat fd/422 mutant wing disc () ordiap1-lacz in a d G13 mutant eye disc (). Anterior is to the left in all discs and arrowheads point to clone borders. Willecke et al. 2of5

3 tubg4 12 ody length D 12 Wing area E 12 Wing length F 12 Wing length / ody length Fig. S3. Uniform overexpression of and produces small flies with even smaller wings. (A) Picture of a wild-type fly. () Picture of a fly that overexpressed and by the tub-gal4 driver. ( F) Quantification of the body length (measured as the distance between the middle of the humeral bristles and the base of the haltere), wing area, wing length, and wing length divided by the body length of flies with the indicated genotypes. Flies with uniform and expression are smaller than wild-type siblings () and have smaller wings (D and E). Such small flies have wings that are comparatively smaller as shown by the reduced wing length to body length ratio compared with wild-type flies (F). Areas and distances were normalized to the wild-type control flies. Measurements were done by using the histogram function of Adobe Photoshop. Willecke et al. 3of5

4 A P(red) : A(green) ratio hhg4 hhg4 hhg4- Fig. S4. Overexpression of and in the posterior compartment produces wings with small posterior compartments. (A) Picture of a wing from a wild-type fly. () Picture of a wing from a fly that overexpressed and in the posterior compartment by the hh-gal4 driver. () Quantification of the ratio of the size of the posterior side (area between veins 4 and the posterior edge in red) and the anterior side that did not express and (area between veins 1 and 3 in green). Flies with overexpression of and in the posterior compartment produced wings that had significantly smaller posterior compartments compared with wild-type controls. Measurements were done using the histogram function of Adobe Photoshop. Willecke et al. 4of5

5 overexpression clone mutant clone activity proximal distal proximal distal Hippo target gene expr. ell polarity Fig. S5. Schematic representation of boundary effects on planar cell polarity and Hippo signaling. (Top) The gradient of activity in tissues with a clone of cells overexpressing (Left) and a ds mutant clone (Right). (Middle) Representation of the pattern of Hippo target gene induction. Target genes are induced in gradients on both sides of the overexpression clone, because clones of cells with abnormal levels of activity create discontinuities in the amount of activity all around clone borders. ds mutant clones also cause induction of Hippo target genes, but only in containing cells, indicating that is required in cells to respond to the boundary signal. (ottom) The direction of cell polarity is toward low levels of activity. ecause clones of cells with abnormal levels of activity change the direction of the gradient only on one side of the clone, cell polarity is reversed only on this side. Green arrows are normal polarity and red arrows depict reversed polarity. In contrast to the induction of Hippo target genes, cell polarity reversals can propagate into ds mutant clones, as was observed in the eye for ommatidial polarity. This indicates that pathways other than signaling also regulate planar cell polarity. Willecke et al. 5of5