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. 2012 May 22;22(10):907–914. doi: 10.1016/j.cub.2012.03.053

Figure 2.

Figure 2

Asymmetry of Ft and Ds Distribution

(A and B) Confocal images of apicolateral junctions in third-instar wing discs near the dorsal hinge. Labeled for (A) Dachs (magenta) and Ds (green) and (B) Dachs (magenta) and Ft (green). Arrowheads point to Dachs and Ds or Ft colocalization on PD cell boundaries.

(C) Images of patches of Ds-EGFP expression in the wing and dorsal eye disc showing Ds-EGFP (green or white) and Arm (magenta). By using a fly strain in which ds is tagged at its endogenous locus with EGFP, we generated patches of Ds-EGFP expression abutting cells expressing untagged Ds but with no change in gene dosage (ds-EGFP FRT40/ ds+ FRT40). Ds-EGFP is enriched at distal cell edges in the wing disc (distal is bottom) and equatorial and posterior cell edges in the eye disc (equator is top and posterior right). Arrowheads point to Ds enrichment and arrows point to reduced Ds.

(D) Twenty-eight hr pupal wing containing a clone overexpressing Ds (Act>stop>GAL4/UAS-ds) labeled for Ds (green) and Dachs (magenta). Arrowheads point to cell boundaries outside the overexpression clone where Dachs and Ds colocalize. Note that as Dachs and Ds asymmetry is lost by this stage of development ([2]; data not shown), the Dachs and Ds asymmetry generated around the clone is due to Ds overexpression. Additionally asymmetric localization is more easily observed in the pupal wing, because cells are larger and more uniform than in wing and eye discs and cell division is no longer occurring.

(E) Ratio of mean fluorescence intensity of Actin, Dachs, Ft, and Ds labeling on PD compared to AP cell boundaries in wing discs close to the dorsal hinge. Error bars show SEM between wing discs (n = 10). A one-way ANOVA test was applied (significance levels [∗∗∗p < 0.001, ∗∗p < 0.01, ∗p < 0.05] between columns linked by bars). Dachs, Ds, and Ft show enrichment on PD boundaries compared to cortical actin. Enrichment of Ft (∗p > 0.05) and Ds (∗∗p < 0.01) is not as strong as Dachs.

(F and G) Mean fluorescence intensity levels of Ds-EGFP staining at cell junctions on the edges of clones in wing (F) and eye discs (G) normalized to WT levels on distal or equatorial cell junctions. Error bars show SEM. One-way ANOVA tests were applied (significant differences linked by bars). In wing discs, Ds-EGFP is significantly higher on distal cell junctions than posterior, anterior, or proximal junctions. In the eye, Ds-EGFP is signficantly higher on equatorial and posterior cell junctions than polar or anterior boundaries.

(H) Mean fluorescence intensity of Dachs (blue), Ds (red), and Arm (yellow) on cell boundaries parallel to UAS-ds overexpression clones in the pupal wing. Intensity was normalized to mean levels on boundaries away from the clone. The cell boundary at the clone edge is labeled 0 (zero). Error bars show SEM between clones (n = 10). A one-way ANOVA test was applied comparing each column with values on cell boundaries away from the clone (column 6). When averaged over ten clones, significant increases in Dachs and Ds were only found around 1–2 cells away from the clone boundary, although in some clones, differences are visible by eye 3–4 cells away; for example, see the clone in (D).

See also Figure S2.