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. 2013 Jan 11;6(3):661–678. doi: 10.1242/dmm.010066

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© 2013. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Share Alike License (http://creativecommons.org/licenses/by-nc-sa/3.0), which permits unrestricted non-commercial use, distribution and reproduction in any medium provided that the original work is properly cited and all further distributions of the work or adaptation are subject to the same Creative Commons License terms.

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Fig. 3. — Reducing levels of Rok in RhoGEF2 + Raf^GOF co-expressing EAD clones suppressed clonal tissue accumulation, differentiation and cell morphology defects, and rescued pupation. Confocal planar sections through the epithelium of day 5 AEL third instar larval EADs. Apical (A,C) and basal (B,D) sections are shown. Mutant tissue was marked by the expression of GFP (green). EADs were stained for Elav (white) and with phalloidin-TRITC for F-actin (white). (A,B) rok^RNAi + RhoGEF2 + Raf^GOF. (C,D) rok^RNAi+ RhoGEF2. Expression of rok^RNAi in RhoGEF2 + Raf^GOF clones resulted in reduced accumulation of undifferentiated clonal tissue compared with RhoGEF2 + Raf^GOF mosaic eye discs (compare arrows, B-Bii and Fig. 2B-Bii) and reduced the high levels of F-actin in the mutant cells (arrowheads in Aiii,Biii compared with Fig. 2Aii,Aiii,Bii,Biii). rok^RNAi in RhoGEF2 clones rescued the reduced clone size, F-actin accumulation and morphology defects of RhoGEF2-alone clones (arrowheads in Cii,Ciii,Dii,Diii compared with supplementary material Fig. S1Aii,Aiii,Bii,Biii). Note that the high levels of F-actin observed in Biii are due to the axonal projections that are present in this basal section. (E) Expression of rok^RNAi in RhoGEF2 + Raf^GOF-expressing cells resulted in an increase in pupation compared with RhoGEF2 + Raf^GOF expression alone. The data was compared by a t-test and error bars represent s.e.m. The significance was P<0.0001. (F) Quantification of F-actin levels in RhoGEF2 + Raf^GOF + rok^RNAi or RhoGEF2 + rok^RNAi clones versus wild-type clones. The data was compared by a t-test and error bars represent s.e.m. The significance was P<0.0004 for RhoGEF2 + rok^RNAi versus RhoGEF2 and P<0.0001 for RhoGEF2 + Raf^GOF + Rho1^RNAi versus RhoGEF2 + Raf^GOF.