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. 2018 Feb 5;46(8):4176–4187. doi: 10.1093/nar/gky071

Figure 2.

Figure 2.

eIF4A interacts with the base of the DLP RNA in the 48S complex. (A) Crosslinking assays using SV-DLP U1 and SFV-DLP U1 mRNAs in the presence of hippuristanol or polyI:C. Samples were treated with 1 μM of hippuristanol or 0.5 ng/μl poly I:C to block eIF4A or to induce eIF2α phosphorylation, respectively, as indicated. Then, P100 fractions were obtained and equivalent cpms were analyzed by SDS-PAGE and autoradiography. The lower panel shows typical eIF2α phosphorylation induced by poly I:C treatment of RRL. (B) Denaturing IP with anti-eIF4A and anti-RPS6 antibodies. 48S complexes were assembled with SV-DLP U1 mRNA, crosslinked and the WRF was denatured and subjected to IP with the indicated antibodies (as described in ‘Materials and Methods’ section).