We could not reliably assess specific marker expression for MFH-MPNST, and MFHSYN given the small number of tumors predicted as these categories

r 5 final washes with PBS, pH 7.2, samples were mounted and observed under the microscope. The image shown is composed from images taken from different fields and assembled using Photoshop. Bar: 15 mm. In quantification of number of fibers per cell is shown. Serial Z-images were obtained per each cell and maximal projections were used to determine the number of fibers per cell. Craven RA, 24195657 Griffiths DJ, Sheldrick KS, Randall RE, Hagan IM, Carr AM. Vectors for the expression of tagged proteins in Schizosaccharomyces pombe. Gene 221: 59-68. Found at: doi:10.1371/journal.pone.0012933.s001 Plant infections, mating assays and germination of teliospores Pathogenic development of wild type and mutant strains was assayed by plant infections of the maize variety Early Golden Bantam as described. For charcoal mating assays, strains were crossed on charcoal-containing complete medium plates and incubated at 22uC. To assay the germination of teliospores, infected plants were incubated for 20 days. Tumors containing spores were dried at room temperature and minced using a mortar and pestle. The tumor material was incubated overnight in a 1.5% copper sulfate. After extensive washing in sterile distilled water, spores were plated on 2% complete medium-containing agar slides and incubated in a moist chamber at 22uC or 34uC. Microscopy Images were obtained either using a Nikon Eclipse 90i fluorescence microscope with a Hamamatsu Orca-ER camera driven by Metamorph or a DeltaVision RT Odanacatib price Restoration Microscopy System with a Coolsnap HQ camera driven by SoftWoRx v.3.5.0 Software. One focal plane images are shown unless otherwise specified. Nikon Eclipse images are shown unless otherwise specified. Images were further processed with Adobe Photoshop CS or Imaris 6.0.1 software. Acknowledgments We thank Prof. Gero Steinberg for the gift of RFP-Tub1 construction as well as for critical reading of the manuscript, Prof. Iain Hagan for the gift of pk constructions and antibodies, and the Centro Andaluz de Biologia del Desarrollo for using their DeltaVision Microscopy System. Supporting Information Author Contributions Conceived and designed the experiments: IAT JPM. Performed the experiments: IAT JPM. Analyzed the data: IAT JPM. Wrote the paper: IAT JPM. 15 September 2010 | Volume 5 | Issue 9 | e12933 Septins in Corn Smut Fungus 16 September 2010 | Volume 5 | Issue 9 | e12933 Septins in Corn Smut Fungus 63. Becht P, Konig J, Feldbrugge M The RNA-binding protein Rrm4 is essential for polarity in Ustilago maydis and shuttles along microtubules. J Cell Sci 119: 4964973. 64. Becht P, Vollmeister E, Feldbrugge M Role for RNA-binding proteins implicated in pathogenic development of Ustilago maydis. Eukaryot Cell 4: 12133. 65. Lindsey R, Cowden S, Hernandez-Rodriguez Y, Momany M Septins AspA and AspC are important for normal development and limit the emergence of new growth foci in the multicellular fungus Aspergillus nidulans. Eukaryot Cell 9: 15563. 66. Tsukuda T, Carleton S, Fotheringham S, Holloman WK Isolation and characterization of an autonomously replicating sequence from Ustilago maydis. Mol Cell Biol 8: 3703709. 67. Straube A, Weber I, Steinberg G A novel mechanism of nuclear envelope break-down in a fungus: nuclear migration strips off the envelope. EMBO J 24: 1674685. 68. Straube A, Enard W, Berner A, Wedlich-Soldner R, Kahmann R, et al. A split motor domain in a cytoplasmic dynein. EMBO J 20: 5091100. 69. Brachmann A, Konig J, Julius C, Feldbrugge M A reverse g

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