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. 2011 Jul 14;39(19):8404–8415. doi: 10.1093/nar/gkr555

Figure 2.

Figure 2.

Proteins associated with LeishIF4E-1 in promastigotes. (A) Pull-down experiments with the SBP-tagged LeishIF4E-1 from L. amazonensis promastigotes was done using affinity purification and further analysis as described in Figure 1A. The antibodies used were directed against LeishIF4E-1, LeishIF4G-3 and LeishIF4A-1. Densitometric analysis showed that the elution fraction contained 24, 0 and 3% of the total LeishIF4E-1, LeishIF4G-3 and LeishIF4A-1, respectively. (B) The yeast two-hybrid assay was performed by co-transfecting wild-type YRG-2 yeast strains with pBD fused to LeishIF4A-1, LeishPABP-1 or LeishIF4E-1 and with pAD fused to LeishIF4G-1 through -6 or to LeishIF4E-1. The cells were cultured and spotted as described in ‘Materials and Methods’ section. Expression of the tested proteins was verified by western blot analysis (Supplementary Figure S7).