Figure 5

Similar phenotypic profiles identified genes in pathways and protein complexes. Cells were stained for F-actin (red), α-tubulin (green) and DNA (blue). Distinct phenotypes were observed with dsRNAs targeting different members of the same functional family (for example, GTPases, in the left panels). (a, b) Phenotypes observed in both cell types. (a) RNAi-induced binucleate cell phenotypes identified genes required for cytokinesis, including Rho1 (encoding a GTPase), pebble (a Rho-GEF) and CG10522 (a predicted citron kinase). Kc₁₆₇ cells are shown. (b) RNAi resulting in loss of actin filaments from the cell cortex identified regulators of actin-filament formation, including Cdc42 (GTPase), enabled (actin-binding protein) and SCAR (actin-binding, Arp2/3 regulator). Kc₁₆₇ cells (shown) also formed microtubule extensions and a polarized cell shape. (c, d) Some phenotypes were unique to one cell type. (c) RNAi resulting in round, non-adherent S2R⁺ cells identified genes required for cell-matrix adhesion, including Roughened (a Rap1 GTPase), Tenascin-major (an adhesion protein with a laminin domain) and myospheroid (β integrin). (d) An RNAi-induced amorphous S2R⁺ cell phenotype identified genes in the mitogen-activated protein (MAP) kinase pathway, including Ras85D (a GTPase), Downstream of raf1 (a MAP kinase kinase, or MEK) and kinase suppressor of Ras (a MAP kinase scaffold protein).