Requirements for eIF4A and eIF2 during translation of Sindbis virus subgenomic mRNA in vertebrate and invertebrate host cells
Article first published online: 20 DEC 2012
© 2012 Blackwell Publishing Ltd
Volume 15, Issue 5, pages 823–840, May 2013
How to Cite
Garcia-Moreno, M., Sanz, M. A., Pelletier, J. and Carrasco, L. (2013), Requirements for eIF4A and eIF2 during translation of Sindbis virus subgenomic mRNA in vertebrate and invertebrate host cells. Cellular Microbiology, 15: 823–840. doi: 10.1111/cmi.12079
- Issue published online: 14 APR 2013
- Article first published online: 20 DEC 2012
- Accepted manuscript online: 27 NOV 2012 06:32PM EST
- Manuscript Accepted: 20 NOV 2012
- Manuscript Revised: 8 NOV 2012
- Manuscript Received: 21 SEP 2012
- DGICYT (Dirección General de Investigación Científica y Técnica. Ministerio de Educación y Ciencia. Spain). Grant Number: BFU2009-07352
- Canadian Cancer Society Research Institute
Fig. S1. Subcellular localization of eEF2 and SV C protein in mock and SV-infected BHK cells. BHK cells were seeded on glass coverslips and mock-infected or infected with SV (5 pfu per cell). At 3.5 or 6.5 hpi, cells were treated for 90 min with 0.5 μM hipp or left untreated. At 5 or 8 hpi, cells were fixed, permeabilized and processed for immunofluorescence using anti-eEF2 (red) and anti-SV C protein (cyan) respectively. Images were acquired on a confocal microscope and subsequently processed with Huygens 4.1 software. Merged images represent the dual visualization of eEF2 and C.
Fig. S2. In vitro translation of SV mRNAs in RRL. Effect of hipp treatment. One hundred nanograms of sgmRNA C+luc or gmRNA SV-Luc mRNAs synthesized in vitro by T7 RNA polymerase were added to RRL in the presence of different concentrations of hipp (0, 0.8 and 1.6 μM) and incubated for 90 min at 30°C. Luc synthesis was estimated by measuring luc activity. The percentage values of hipp-treated samples relative to their respective untreated samples are represented in the figure and are the mean ± SD of three representative experiments performed in triplicate.
Table S1. Nomenclature of plasmids, replicons, sgmRNAs and mRNAs.
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