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. 2020 Mar 30;48(8):4405–4417. doi: 10.1093/nar/gkaa173

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© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Association of Leish4E-IP2 with different LeishIF4Es. The association between Leish4E-IP2 and LeishIF4E-1 (4E1), LeishIF4E-4 (4E4) (A) and LeishIF4E-3 (4E3) (B) were monitored by pull-down experiments of mid-log L. amazonensis promastigotes over-expressing SBP-tagged LeishIF4E-IP2. The cells were lysed and affinity-purified over streptavidin-Sepharose beads. The beads were washed and further eluted with biotin. Aliquots from the soluble extract (S, 5%), the flow-through fraction (FT, 5%), the final wash (W, 50%) and eluted proteins (E, 50%) were separated by 10% SDS–PAGE and subjected to western blot analysis using specific antibodies. The antibodies used were raised against LeishIF4E-1, LeishIF4E-4 (A), LeishIF4E-3 and LeishIF4A (as a negative control) (B) along with anti-SBP antibodies that were used to highlight the SBP-tagged Leish4E-IP2. (C) Representation of the densitometry analysis of each quantity of LeishIF4E co-eluted by Leish4E-IP2.