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. 2013 Jul 11;9(7):e1003598. doi: 10.1371/journal.pgen.1003598

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© 2013 Baumgartner 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–G) Scanning electron micrographs of adult control (A), lig^{RNAi II} (B), FMR1^D113M (C), lig^{RNAi II} FMR1^D113M (D), rin² (E), lig^{RNAi II} rin² (F) eyes generated by eyFLP Actin-Flp out-Gal4/FRT-mediated mitotic recombination from flies grown at 25% yeast content (A–F). Scale bar represents 100 µm. (G) Statistical analyses as described in Figure 1D: control (714±30 and 720±30), lig^{RNAi II} expressing (738±32 and 706±23), FMR1^D113M mutant (737±10 and 729±34), lig^{RNAi II} expressing FMR1^D113M mutant (775±36; p = 0.0051 and 806±23; p = 0.00046), rin² mutant (767±34 and 760±40), lig^{RNAi II} expressing rin² mutant (821±45; p = 0.03 and 817±69; p = 0.082) eyes from flies raised on 25% yeast-containing food. (H–J) Scanning electron micrographs of adult control (H), lig^{RNAi II} (I) and lig^{RNAi II} Capr² (J) eyes generated by eyFLP Actin-Flp out-Gal4/FRT Minute-mediated mitotic recombination from flies grown on 25% yeast-containing food. (K–M'') Negatively marked 72 h old lig¹ mutant clones (induced with the hsFLP/FRT system) in eye imaginal discs of third instar larvae (K', L' and M'). FMR1 levels (visualized by immunostaining) remain unchanged (K''). Capr levels (visualized by immunostaining) are slightly increased in lig mutant clones (L''). Rin-Cherry levels expressed from the Grin^Cherry transgene are autonomously decreased in the lig¹ mutant clones (M''). Imaginal discs are stained with DAPI (blue) to visualize the DNA. Scale bar represents 25 µm. (N) S2 cells transfected with GFP-Lig and Grin^Cherry have increased levels of Rin-Cherry in comparison to S2 cells overexpressing GFP and Grin^Cherry. S2 cells overexpressing GFP-Lig upregulate a transcriptional reporter consisting of the rin promoter followed by a Cherry coding sequence and the 3′ UTR of rin. Conversely, a translational reporter consisting of an ubi promoter followed by the 5′ UTR of rin, a Cherry protein coding sequence and the 3′ UTR of rin is not affected. Genotypes: (A) y w eyFLP, Act>CD2>Gal4/y w; FRT82 cl w⁺/FRT82 (B) y w eyFLP, Act>CD2>Gal4/y w; UAS-lig^{RNAi II} [51D]/+; FRT82 cl w⁺/+ (C) y w eyFLP, Act>CD2>Gal4/y w; FRT82 cl w⁺/FRT82 FMR1^D113M (D) y w eyFLP, Act>CD2>Gal4/y w; UAS-lig^{RNAi II} [51D]/+; FRT82 cl w⁺/FRT82 FMR1^D113M (E) y w eyFLP, Act>CD2>Gal4/y w; FRT82 cl w⁺/FRT82 rin² (F) y w eyFLP, Act>CD2>Gal4/y w; UAS-lig^{RNAi II} [51D]/+; FRT82 cl w⁺/FRT82 rin² (H) y w eyFLP, Act>CD2>Gal4/y w; UAS-CG1315^RNAi (control)/+; M(3)RpS17⁴ FRT80/+ (I) y w eyFLP, Act>CD2>Gal4/y w; UAS-lig^{RNAi II} [51D]/+; M(3)RpS17⁴ FRT80/+ (J) y w eyFLP, Act>CD2>Gal4/y w; UAS-lig^{RNAi II} [51D]/+; M(3)RpS17⁴ FRT80/Capr² FRT80 (K, L) y w hsFLP/y w; FRT42 ubiGFP/FRT42 lig¹ (M) y w hsFLP/y w; FRT42 ubiGFP/FRT42 lig¹; Grin^Cherry [86Fb]/+.