These authors contributed equally to this work.
Identification of novel Drosophila centromere-associated proteins
Version of Record online: 14 JUL 2014
© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Special Issue: Proteomics in Chromatin Biology and Epigenetics
Volume 14, Issue 19, pages 2167–2178, October 2014
How to Cite
Barth, T. K., Schade, G. O. M., Schmidt, A., Vetter, I., Wirth, M., Heun, P., Thomae, A. W. and Imhof, A. (2014), Identification of novel Drosophila centromere-associated proteins. Proteomics, 14: 2167–2178. doi: 10.1002/pmic.201400052
- Issue online: 2 OCT 2014
- Version of Record online: 14 JUL 2014
- Accepted manuscript online: 19 MAY 2014 06:33PM EST
- Manuscript Accepted: 15 MAY 2014
- Manuscript Revised: 3 APR 2014
- Manuscript Received: 16 FEB 2014
- German Research Council. Grant Number: IM 23/9—1
- European Union. Grant Number: 257082
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Figure S1. (A) Immunolocalizations on mitotic chromosome spreads demonstrate centromeric association of CG3548, but not CG2051 and CG14480, during mitosis. Green: GFP-tagged construct, red: CID, blue: DAPI. Scale bars represent 3 μm. Pictures were deconvolved and maximum intensity projections are shown. (B) CG11076 localizes to the nucleolus as indicated by colocalization with Fibrillarin. Green: GFP-tagged construct, red: Fibrillarin, blue: DAPI. Scale bars represent 3 μm. Pictures were deconvolved and a single optical section is shown.
Figure S2. CID-GFP chromatin copurifying factors that show nuclear localization. Immunolocalization of GFP-tagged candidate proteins in stable cell lines (CG11076) or transient transfections (others). Centromeres were visualized by anti-CID antibody staining. Line profiles indicate the distribution of signals in the different channels. Blue: DAPI, green: epitope-tagged candidate protein, red: CID. Scale bars represent 3 μm. Pictures were deconvolved and a single optical section is shown.
Figure S3. CID-GFP chromatin copurifying factors that show localization outside the nucleus. Immunolocalization of GFP-tagged candidate proteins in stable cell lines (CG1091) or transient transfections of epitope-tagged proteins (others). Centromeres were visualized by anti-CID antibody staining. Blue: DAPI, green: epitope-tagged candidate protein, red: CID. Scale bars represent 3 μm. Pictures were deconvolved and a single optical section is shown.
Table S1. MaxQuant result of 1871 detected proteins in nine purifications. Indicated are protein names, razor and unique peptides, molecular weight, PEP (posterior error probability), intensities and iBAQ values. The data were sorted in descending order according to the following parameters: i) log2-fold enrichment over both controls of more than four (column AC); ii) identified in at least two CID-chromatin purifications (column AE); iii) sum of iBAQ values in CID-chromatin purifications exceeding 90% of the sum of all iBAQ values (column AI); iv) average iBAQ value in CID-chromatin purification (column AF).
Table S2. 86 candidates displaying more than 4-fold enrichment in CID-GFP chromatin purifications over both control conditions and five additionally tested candidates specific for CID-GFP purifications (see Material & Methods section). Indicated are protein names, log2 values of average enrichments in three replicates with standard deviation and p-values, experimental evidence from this work, available expression constructs.
Table S3. Classification of immunolocalizations. An overview of different cellular localizations of tested factors with representative pictures is given.
Table S4. Raw data of CID-GFP quantification for box plots (corresponding to figure 3C).
Table S5: RNAi primers used to knockdown expression of selected CID interactors.
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