Fig. 4. Expression of miR-2 family mediates tissue growth of Hippo pathway.(A) Photomicrographs of adult wings of the indicated genotype. Expression of.

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Fig. 4. Expression of miR-2 family mediates tissue growth of Hippo pathway.(A) Photomicrographs of adult wings of the indicated genotype. Expression of miR-2 family mediates tissue growth of Hippo pathway.(A) Photomicrographs of adult wings of the indicated genotype. Left panels: ptc-Gal4 control flies. Right panels ptc-Gal4 driving expression of a UAS-ykiRNAi transgene to reduce yki mRNA levels in the ptc-Gal4 expression domain. Upper panels: control flies without expressing any other transgene. Lower panels: flies expressing UAS-miR-2a/2b cluster transgene to increase miR-2a and 2b levels. Histogram at right shows quantification of the effects on growth of the ptc-Gal4 expression domain. Error bars represent standard deviation from measurement of at least 8 wings for each genotype. *** indicates statistically significant increase in the width of the ptc-Gal4 expression domain when miR-2 cluster level was increased (P<0.001). (B) Photomicrographs of adult wings of the indicated genotype, as in panel A. Lower panels: flies expressing a UAS-miR-11 transgene to increase miR-11 level in the ptc-Gal4 domain. Histogram shows quantification of the effects of the UAS-miR-11 transgene alone and together with UAS-ykiRNAi. Error bars represent standard deviation from at least 6 wings for each genotype. *** indicates statistically significant increase in the width of the ptc-Gal4 expression domain when miR-11 level was increased (P<0.001). As a member of the miR-2 seed family, miR-11 is expected to regulate the same targets as miR-2. Regulation of reaper by miR-11 has been confirmed in the embryo by Leaman et al. and Ge et al. (Leaman et al., 2005; Ge et al., 2012). (C) Histogram showing the levels of miR-2 and miR-13 measured by quantitative RT-PCR. Wandering 3rd instar larvae expressed UAS-ykiRNAi under ubiquitous tubulin-Gal4 control. Control (white bars) expressed tubulin-Gal4 without the UAS-RNAi transgene. Total RNA from whole larvae was extracted and normalized for cDNA synthesis. RNA levels were normalized to small RNA U14. Error bars represent standard deviation from 3 independent experiments. Student's t-test for miR-2 vs U14: *P<0.05, **P<0.01. (D) Yki acts at multiple levels to control apoptosis. Arrows indicate activation, whereas bar-ended lines indicate inhibitory interactions. Wei Zhang, and Stephen M. Cohen Biology Open 2013;bio.20134317 © 2013. Published by The Company of Biologists Ltd