Research
Identification of secondary targets of N-containing bisphosphonates in mammalian cells via parallel competition analysis of the barcoded yeast deletion collection
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* Corresponding author: Daniela Delneri d.delneri@manchester.ac.uk
- † Equal contributors
1 Department of Biomedical Sciences and Technologies, University of Udine, Piazzale Kolbe, 33100, Udine, Italy
2 Faculty of Life Science, University of Manchester, Oxford Road, M13 9PT, Manchester, UK
3 School of Biological Sciences, Institute of Evolutionary Biology, King's Buildings, West Mains Road, Edinburgh EH9 3JT, UK
4 The Center for the Study of Metabolic Bone Diseases, via Vittorio Veneto, 34170, Gorizia, Italy
5 Department of Medical and Morphological Research, University of Udine, Piazzale Kolbe, 33100, Udine, Italy
Genome Biology 2009, 10:R93 doi:10.1186/gb-2009-10-9-r93
Published: 10 September 2009Additional files
Additional data file 1:
Figure S1: wild-type S. cerevisiae responds to the N-BPs in a dose-dependent manner. All three drugs are able to inhibit growth and equimolar doses of each drug display a different degree of toxicity, with RIS and IBA being the most powerful. Representative growth curves of wild-type S. cerevisiae BY4743, grown in the presence of increasing concentrations of the indicated N-BPs for 20 h. Yeast growth was monitored using an OD reader with measurements every 5 minutes. Figure S2: N-BP sensitivity profiling for the significant strains. Each bar in the graph represents a specific strain; the more negative the value of the bar, the greater the rate of diminution of that strain from the pool. Figure S3: FACS analysis of the MCF-7 cell line treated with N-BPs. Percentage of cells in G0/G1, S and G2/M phases (A) after the exclusion of the sub-G1 population (dead cells), which was analyzed separately (B). Figure S4: effect of N-BPs on cell migration. MCF-7 cells were treated with or without 100 μM N-BP as described in Materials and methods and observed by time-lapse microscopy for the next 48 h, taking phase-contrast photographs every 4 h. The horizontal bars represent the limit of the slit performed on the cell monolayer at the start of the experiment. Five measurements per well were taken; the figure shows a representative experiment at 24 h and 48 h. Original magnification 200×.
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Additional data file 2:
QD strains are shown in red.
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Additional data file 3:
Haploproficient strains (q < 0.01).
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Additional data file 4:
Haploinsufficient and QD strains after removal of bad tags.
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Additional data file 5:
Haploproficient strains after removal of bad tags.
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Additional data file 6:
Growth data and two-way ANOVA of the wild-type (WT) strain and the hemizygote mutants DBF4 (A) and ALF1 (B) in the presence and absence of the drug ibandronate (IBA).
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