FIG 2.

Replication of decapping enzyme mutant and E3 deletion virus in HAP1 and A549 KO cells. (A) Absence of RNase L and PKR proteins in HAP1 KO cells. Cell lysates from HAP1 control, RNase L KO, PKR KO, and DKO cells were analyzed by Western blotting using mouse polyclonal antibody to RNase L and rabbit MAb to PKR. Antibody to actin was used as a loading control. (B) One-step virus replication. HAP1 control, RNase L KO, PKR KO, and DKO cell monolayers in 12-well plates were infected in triplicate with 5 PFU/cell of purified vD10rev, vD9muD10mu, or vΔE3L and harvested at 3 and 24 h. Virus titers were determined by plaque assay in BHK-21 cells. The 3-h titers represent input virus. (C and D) Procedures were the same as those described for panels A and B except that A549 control and KO cells were used. Each bar represents the standard deviation determined from three replicate infections.