10.1371/journal.pbio.1001199.g004 Christopher A. Maxwell Christopher A. Maxwell Javier Benítez Javier Benítez Laia Gómez-Baldó Laia Gómez-Baldó Ana Osorio Ana Osorio Núria Bonifaci Núria Bonifaci Ricardo Fernández-Ramires Ricardo Fernández-Ramires Sylvain V. Costes Sylvain V. Costes Elisabet Guinó Elisabet Guinó Helen Chen Helen Chen Gareth J. R. Evans Gareth J. R. Evans Pooja Mohan Pooja Mohan Isabel Català Isabel Català Anna Petit Anna Petit Helena Aguilar Helena Aguilar Alberto Villanueva Alberto Villanueva Alvaro Aytes Alvaro Aytes Jordi Serra-Musach Jordi Serra-Musach Gad Rennert Gad Rennert Flavio Lejbkowicz Flavio Lejbkowicz Paolo Peterlongo Paolo Peterlongo Siranoush Manoukian Siranoush Manoukian Bernard Peissel Bernard Peissel Carla B. Ripamonti Carla B. Ripamonti Bernardo Bonanni Bernardo Bonanni Alessandra Viel Alessandra Viel Anna Allavena Anna Allavena Loris Bernard Loris Bernard Paolo Radice Paolo Radice Eitan Friedman Eitan Friedman Bella Kaufman Bella Kaufman Yael Laitman Yael Laitman Maya Dubrovsky Maya Dubrovsky Roni Milgrom Roni Milgrom Anna Jakubowska Anna Jakubowska Cezary Cybulski Cezary Cybulski Bohdan Gorski Bohdan Gorski Katarzyna Jaworska Katarzyna Jaworska Katarzyna Durda Katarzyna Durda Grzegorz Sukiennicki Grzegorz Sukiennicki Jan Lubiński Jan Lubiński Yin Yao Shugart Yin Yao Shugart Susan M. Domchek Susan M. Domchek Richard Letrero Richard Letrero Barbara L. Weber Barbara L. Weber Frans B. L. Hogervorst Frans B. L. Hogervorst Matti A. Rookus Matti A. Rookus J. Margriet Collee J. Margriet Collee Peter Devilee Peter Devilee Marjolijn J. Ligtenberg Marjolijn J. Ligtenberg Rob B. van der Luijt Rob B. van der Luijt Cora M. Aalfs Cora M. Aalfs Quinten Waisfisz Quinten Waisfisz Juul Wijnen Juul Wijnen Cornelis E. P. van Roozendaal Cornelis E. P. van Roozendaal Douglas F. Easton Douglas F. Easton Susan Peock Susan Peock Margaret Cook Margaret Cook Clare Oliver Clare Oliver Debra Frost Debra Frost Patricia Harrington Patricia Harrington D. Gareth Evans D. Gareth Evans Fiona Lalloo Fiona Lalloo Rosalind Eeles Rosalind Eeles Louise Izatt Louise Izatt Carol Chu Carol Chu Diana Eccles Diana Eccles Fiona Douglas Fiona Douglas Carole Brewer Carole Brewer Heli Nevanlinna Heli Nevanlinna Tuomas Heikkinen Tuomas Heikkinen Fergus J. Couch Fergus J. Couch Noralane M. Lindor Noralane M. Lindor Xianshu Wang Xianshu Wang Andrew K. Godwin Andrew K. Godwin Maria A. Caligo Maria A. Caligo Grazia Lombardi Grazia Lombardi Niklas Loman Niklas Loman Per Karlsson Per Karlsson Hans Ehrencrona Hans Ehrencrona Anna von Wachenfeldt Anna von Wachenfeldt Rosa Bjork Barkardottir Rosa Bjork Barkardottir Ute Hamann Ute Hamann Muhammad U. Rashid Muhammad U. Rashid Adriana Lasa Adriana Lasa Trinidad Caldés Trinidad Caldés Raquel Andrés Raquel Andrés Michael Schmitt Michael Schmitt Volker Assmann Volker Assmann Kristen Stevens Kristen Stevens Kenneth Offit Kenneth Offit João Curado João Curado Hagen Tilgner Hagen Tilgner Roderic Guigó Roderic Guigó Gemma Aiza Gemma Aiza Joan Brunet Joan Brunet Joan Castellsagué Joan Castellsagué Griselda Martrat Griselda Martrat Ander Urruticoechea Ander Urruticoechea Ignacio Blanco Ignacio Blanco Laima Tihomirova Laima Tihomirova David E. Goldgar David E. Goldgar Saundra Buys Saundra Buys Esther M. John Esther M. John Alexander Miron Alexander Miron Melissa Southey Melissa Southey Mary B. Daly Mary B. Daly Rita K. Schmutzler Rita K. Schmutzler Barbara Wappenschmidt Barbara Wappenschmidt Alfons Meindl Alfons Meindl Norbert Arnold Norbert Arnold Helmut Deissler Helmut Deissler Raymonda Varon-Mateeva Raymonda Varon-Mateeva Christian Sutter Christian Sutter Dieter Niederacher Dieter Niederacher Evgeny Imyamitov Evgeny Imyamitov Olga M. Sinilnikova Olga M. Sinilnikova Dominique Stoppa-Lyonne Dominique Stoppa-Lyonne Sylvie Mazoyer Sylvie Mazoyer Carole Verny-Pierre Carole Verny-Pierre Laurent Castera Laurent Castera Antoine de Pauw Antoine de Pauw Yves-Jean Bignon Yves-Jean Bignon Nancy Uhrhammer Nancy Uhrhammer Jean-Philippe Peyrat Jean-Philippe Peyrat Philippe Vennin Philippe Vennin Sandra Fert Ferrer Sandra Fert Ferrer Marie-Agnès Collonge-Rame Marie-Agnès Collonge-Rame Isabelle Mortemousque Isabelle Mortemousque Amanda B. Spurdle Amanda B. Spurdle Jonathan Beesley Jonathan Beesley Xiaoqing Chen Xiaoqing Chen Sue Healey Sue Healey Mary Helen Barcellos-Hoff Mary Helen Barcellos-Hoff Marc Vidal Marc Vidal Stephen B. Gruber Stephen B. Gruber Conxi Lázaro Conxi Lázaro Gabriel Capellá Gabriel Capellá Lesley McGuffog Lesley McGuffog Katherine L. Nathanson Katherine L. Nathanson Antonis C. Antoniou Antonis C. Antoniou Georgia Chenevix-Trench Georgia Chenevix-Trench Markus C. Fleisch Markus C. Fleisch Víctor Moreno Víctor Moreno Miguel Angel Pujana Miguel Angel Pujana Genetic interactions influencing epithelial apicobasal polarization. Public Library of Science 2011 interactions influencing epithelial apicobasal 2011-11-15 01:14:06 Figure https://plos.figshare.com/articles/figure/_Genetic_interactions_influencing_epithelial_apicobasal_polarization_/384446 <p>(A) shRNA-mediated depletion of centrosome components impairs polarization. Representative bright-field images are shown for results of untreated and control vector pLKO.1, shRNA-<i>AURKA</i>, shRNA-<i>BRCA1</i>, shRNA-<i>HMMR</i>, or shRNA-<i>TPX2</i> transduced cultures of MCF10A cells in rBM. Magnification is equivalent for all images and scale bars represent 20 µm. (B) Acini architecture was quantified from bright-field images of cultures treated as described above. For comparison between experiments, all values were normalized to untreated cultures within experiments and differences assessed statistically relative to pLKO.1. Shape factor values for single cells, or small clusters, are not plotted. The graph shows the results of at least four independent experiments. For all graphs, asterisks and circles indicate significant differences (two-sided <i>t</i> test <i>p</i><0.05 and <i>p</i><0.005, respectively) from controls (pLKO.1). (C) Representative bright-field images of acini from concurrent depletions (shRNA-mediated) as indicated. (D) <i>AURKA-HMMR</i> interact in the regulation of polarization: <i>HMMR</i> depletion rescues the abnormality seen in the shRNA-<i>AURKA</i> assay. Graph shows the results of three independent experiments. (E) Quantification of acini per well confirms the genetic interaction between <i>AURKA</i> and <i>HMMR</i>. Graph shows the results of duplicate experiments. (F) <i>TPX2</i> depletion is suppressive to abnormalities caused by shRNA-<i>BRCA1</i> and shRNA-<i>HMMR</i>. Graph shows the results of at least three independent experiments. (G) Prior to the shRNA assays, published data proposed the hypothesis of a signaling pathway from TPX2 to RHAMM regulating polarization; degradation of the microtubule-associated factor RHAMM, through BRCA1, was predicted as key to polarization. However, several observations from the single and concurrent depletion assays (depleted proteins are indicated in grey font) diverged from the expected results (divergent observations are italicized). RHAMM depletion impaired polarization in a manner that was rescued by concurrent depletion of AURKA or TPX2, but not BRCA1. On the other hand, concurrent depletion of BRCA1 and TPX2 revealed normal acini.</p>