Figure 6.

Ψ-containing mRNA does not pull-down PKR. Biotinylated in vitro transcribed unmodified or Ψ-containing RNAs were incubated with HEK293T cell lysates for 2 h. The RNA and bound proteins were pulled down using streptavidin-agarose beads. An aliquot of lysate that was incubated only with beads but without RNA (−) was also processed. Aliquots of pull-down proteins as well as the supernatants were separated by SDS–PAGE. PKR and PABP were detected by western blotting. Relative band densities of PKR divided by PABP compared to unmodified RNA are presented below each gel lane.