FIG. 2.

Expression of poliovirus 2A protease in 293-ECR-2AI2A cells promote cleavage of eIF4GI to fragments identical to those observed in poliovirus-infected cells. (A) 293-ECR-2A and 293-ECR-2AI2A cells were treated with MurA (2.5 μM). At the indicated times, the cells were harvested and portions of cellular extracts containing 60 μg of protein were fractionated by electrophoresis on an SDS–8% polyacrylamide gel. The fractionated material was transferred to a nitrocellulose membrane which was then probed with an antibody raised against the C-terminal fragment of eIF4GI. (B) 293-EcR and 293-ECR-2AI2A were treated with MurA (2 μM) or infected with poliovirus (PV) at an MOI of 100 PFU/cell as previously described (13). At the indicated times, cells were harvested and cellular extracts were fractionated by electrophoresis on an SDS–6% polyacrylamide gel and probed with an antibody raised against the N- and C-terminal fragments of eIF4GI. The N (Cp-N)- and C (Cp-C)-terminal cleavage products of eIF4GI are indicated.