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. 2009 Jun 29;185(7):1181–1194. doi: 10.1083/jcb.200904161

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© 2009 Patel et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 7. — nero regulates eIF5A levels in vivo and Nero's DOHH activity is required for nero function. (A) Synchronized larvae 72 h after egg hatching of four different genotypes (from left to right): y w, nero¹, eIF5A^P01296, and eIF5A^P01296; nero¹. The growth of eIF5A^P01296; nero¹ double mutant larvae appears to be more impaired than either eIF5A^P01296 or nero¹ homozygous mutant larvae. (B) Overexpression of eIF5A in nero mutant clones fails to rescue bristle growth defects associated with nero mutant clones (genotype: y w hs-FLP tub-GAL4 UAS-GFP-6xMYC-NLS/+; UAS-eIF5A/+; FRT82B nero¹/FRT82B hsp70-CD2 y⁺ tub-GAL80). (C and D) eIF5A levels are dramatically up-regulated in nero mutant clones in the wing imaginal disc. nero mutant clones are marked by the absence of GFP (green; genotype: y w hs-FLP; FRT82B nero¹/FRT82B Ubi-GFP). White lines mark the clonal boundary. WT, wild type. (E) Overexpression of hDOHH in nero mutant clones rescues bristle size defects (genotype: y w hs-FLP tub-GAL4 UAS-GFP-6xMYC-NLS/+; UAS-hDOHH/+; FRT82B nero¹/FRT82B hsp70-CD2 y⁺ tub-GAL80). (F–I) Overexpression of mutated forms of hDOHH in nero mutant clones fails to rescue bristle size defects. Genotypes are essentially identical to E except mutated forms of hDOHH are expressed under the UAS regulation. (F) Genotype: y w hs-FLP tub-GAL4 UAS-GFP-6xMYC-NLS/+; UAS-DOHH H56A/+; FRT82B nero¹/FRT82B hsp70-CD2 y⁺ tub-GAL80. (G) Genotype: y w hs-FLP tub-GAL4 UAS-GFP-6xMYC-NLS/+; UAS-hDOHH H89A/+; FRT82B nero¹/FRT82B hsp70-CD2 y⁺ tub-GAL80. (H) Genotype: y w hs-FLP tub-GAL4 UAS-GFP-6xMYC-NLS/+; UAS-hDOHH H208A/+; FRT82B nero¹/FRT82B hsp70-CD2 y⁺ tub-GAL80. (I) Genotype: y w hs-FLP tub-GAL4 UAS-GFP-6xMYC-NLS/+; UAS-hDOHH H240A/+; FRT82B nero¹/FRT82B hsp70-CD2 y⁺ tub-GAL80. (B and E–I) Mutant bristles are marked by the recessive yellow mutation and appear light brown.