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. 2011 Apr 1;145(1):79–91. doi: 10.1016/j.cell.2011.02.047

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© 2011 ELL & Excerpta Medica.

Open Access under CC BY 3.0 license

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Drosophila iRhom Inhibits EGFR Signaling

(A) The rough eye caused by sevenless-driven Rhomboid-1 overexpression (sev-rho-1/+) is strongly enhanced by halving the dose of iRhom (iRhom^KO1/+; sev-rho-1/+).

(B) Loss of iRhom suppressed the rough eyes caused by reduction of EGFR signaling mediated by UAS-driven: dominant negative Egfr (U-DN-Egfr); argos (U-argos); and sprouty (U-sty). The driver in all cases was the eye-specific GMR-Gal4.

(C) Expression of UAS-iRhom under the control of GMR-Gal4 suppressed the rough eye caused by sev-rhomboid-1 expression. This was particularly obvious at the anterior edge of the eye indicated by white arrowheads.

(D) Genetic interactions in the wing. Examples of wing phenotypes and severity levels indicated by wild-type (WT), mild (+), and severe (++) are shown. Wings of flies that were heterozygous for argos (argos^lΔ7/+) or iRhom (iRhom^KO1/+) were wild-type; 6.4% (± 6.4%) of wings that were heterozygous for sprouty (sty^S73/+) showed mild extra vein phenotypes (black arrowheads), typical of slight EGFR hyperactivation. A transheterozygous combination of sty^S73 and argos^lΔ7 enhanced the extra vein phenotype (74.5% ± 6.4% with mild extra veins). Similarly, halving iRhom in combination with sty^S73 (iRhom^KO1/+; sty^S73/+) enhanced the phenotype (62.5% ± 11.6% mild; 3.65% ± 5.4% severe). iRhom^KO1/+; sty^S73/ argos^lΔ7 flies showed a further increase in penetrance and strength of the extra vein phenotype (87.5% ± 2.3% mild; 10.3% ± 3.8% severe). All flies for this experiment were grown at 29°C. Error bars represent standard deviations of three to four independent experiments (n = 50 per experiment).

(E) iRhom^KO1/KO1 flies (n = 44) showed increased daytime sleep compared to controls (elav-GAL4, n = 37). Inhibition of EGFR signaling in the nervous system by expression of RNAi constructs against spitz (elav-GAL4; iRhom^KO1/KO1; UAS-spi^RNAi, n = 27) and Egfr (elav-GAL4; iRhom^KO1/KO1; UAS-Egfr^RNAi, n = 36) significantly (p < 0.0001) suppressed the iRhom^KO1/KO1 sleep phenotype. Error bars represent mean ± SEM.

See also Table S1.