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. 2012 Feb 23;8(2):e1002503. doi: 10.1371/journal.pgen.1002503

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Avanesov et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Wild type wing showing the positions of the first through fifth longitudinal veins (L1–L5) and the position of the A/P compartment boundary (dashed line). (B) Reduced L3–L4 spacing in shf mutant wing (B) is not improved upon expression of UAS-shfΔWIF (B′). (C, D) Posterior, en-Gal4-driven expression of UAS-WIF^wif1-EGF^shf (C) or UAS-WIF^shf-EGF^wif1 (D) in shf^x33/Y wings. UAS-WIF^wif1-EGF^shf strongly improved and UAS-WIF^shf-EGF^wif1 weakly improved L3–L4 spacing. (E) Comparison of L3–L4 spacing in wild type, shf², and shf^x33/Y with en-Gal4-driven UAS-construct expression. To compensate for differences in overall wing size, we normalized the L3–L4 distance to the distance between the anterior and posterior margins (red bars). In all but one case we presented the experimental normalized L3–L4 distances as percentages of the wild type normalized distances. However, since expression of UAS-WIF^wif1-EGF^shf reduced the size of the posterior compartment, and thus the distance between the anterior and posterior margins, we compared the normalized L3–L4 distance in shf^x33 UAS-WIF^wif1-EGF^shf wings to the normalized L3–L4 distance in non-shf UAS-WIF^wif1-EGF^shf siblings (n = 31). Bars denote standard deviation. Two-tailed Student's t test showed no significant differences between UAS-WIF^shf-EGF^wif1 and UAS-shfΔEGF, or between shf² and shf², UAS-shfΔWIF. Differences between the other conditions were significant (p<0.0001). (F–I) Anti-Ptc staining in wild type (F), shf^x33/Y (G), and shf^x33/Y wing discs with ap-Gal4-driven expression of UAS-WIF^wif1-EGF^shf (H) or UAS-WIF^shf-EGF^wif1 (I). The improvement in the width of Ptc expression in shf^x33 discs was stronger after expression of UAS-WIF^wif1-EGF^shf than UAS-WIF^shf-EGF^wif1. (J, K) shf^x33/Y wing discs with ap-Gal4-driven expression of UAS-hh-GFP and UAS-WIF^wif1-EGF^shf (J) or UAS-hh-GFP and UAS-WIF^shf-EGF^wif1 (K). UAS-WIF^wif1-EGF^shf strongly improved the ventral accumulation of Hh-GFP, while the improvement with UAS-WIF^shf-EGF^wif1 was more modest (quantifications in Figure S9).