Supplementary MaterialsSupplementary Figure S1 embj0034-0940-sd1. as well as the downstream kinase Warts (LATS1/2 in mammals), which works to phosphorylate and inhibit the transcriptional activator Yorkie (Yki; YAP/TAZ in mammals) (Harvey wing or attention epithelia, mutation of is enough to cause gentle cells overgrowth, but dual mutants result in a stronger overgrowth phenotype, much like or mutants (Hamaratoglu consequently cause a gentle overgrowth phenotype in wing and attention epithelia (Chen or dual mutants result in a solid double mutants highly influence polarisation of Crb within the ovarian follicular epithelium and polarisation from the actin cytoskeleton for boundary cell migration, features that are 3rd party of nuclear signalling via Yki (Fletcher RNAi display within the wing for book genes controlling cells development (M. Campos & B. J. Thompson, manuscript in preparation). In this screen, we identified the apical Spectrin cytoskeleton components -Spectrin (-Spec) and -heavy Spectrin (HSpec)also known as Karst (Kst)as producing moderate wing and eye overgrowth phenotypes, similar to RNAi knock-down of Crb (Fig?(Fig1A1ACF and Supplementary Figs S1 and S2). Spectrins are large cytoskeletal proteins that form hexagonal IRAK inhibitor 1 networks at the intracellular surface of the plasma membrane in all animal cells and have been reported to have mechanosensory properties (Bennett & Baines, 2001; Johnson epithelia, with dimers of -?and H-Spec/Kst localising to the apical domain and dimers of -?and -Spec localising to the basolateral domain (Thomas & Kiehart, 1994; Lee and mutants are known to genetically interact with null mutant in the eye (Fig?(Fig1I1ICR). Open in a separate window Figure 1 The Spectrin cytoskeleton restricts tissue growth in the eye and wing A-O UAS.RNAi lines were driven with for expression during eye development or for expression during wing development. (A, B) Control adult eye (A) and wing (B). (C, D) RNAi results in overgrowth of the eye (C) and wing (D). (E, F) RNAi results in overgrowth of the eye (E) IRAK inhibitor 1 and wing (F). (G, H) RNAi does not affect eye size (G) or wing size (H). (I, J) RNAi results in overgrowth of the eye (I) and wing (J). (K, L) double RNAi results in stronger overgrowth of the eye (K) and wing (L). (M, N) double RNAi results in stronger overgrowth of?the eye (M) and wing (N). (O) Quantification of female wing sizes by pixel area, 5 wings per genotype were measured. Error bars show standard Rabbit Polyclonal to LFNG deviation. P-RThe eyeless FLP MARCM system was used to generate clonally mutant fly eyes. mutant eyes (Q) overgrow slightly compared to controls (P), while mutant eyes expressing RNAi (R) overgrow strongly IRAK inhibitor 1 compared to controls IRAK inhibitor 1 (P). Data information: Scale bars, 250?m. Despite previous reports that apical H-Spec/Kst interacts physically with Crb, genetic analysis of mutants indicated that it is dispensable for polarisation of Crb and for epithelial polarity in general (Thomas (Fig?(Fig2A2ACF). We also examined the expression of the key Hippo reporter gene, RNAi in the posterior compartment with RNAi exhibit a slightly elevated level of expression in the posterior compartment (Fig?(Fig2G2G and IRAK inhibitor 1 ?andH).H). This elevation of expression is similar in magnitude to that caused by RNAi and becomes stronger in double RNAi wing discs, similar to RNAi (Fig?(Fig2I2ICK). These total results show that apical Spectrins regulate Yki activity within the wing and eye. They display that Spectrins work in parallel with Kibra also, very much the same as Former mate (Baumgartner or each result in a more powerful phenotype compared to the single mutants only (Baumgartner mutant pupal retina showing extra interommatidial cells. (C) Pupal retina expressing.