FIG 4.

Broad inhibition of host protein synthesis by the TM domain. (A and B) HEK-293T and IPI-2I cells were transfected with one of the following plasmids: PCAGGS, E66L, E66L(13–34), E66L (1–12), or E66L(35–50) (2.5 μg). The cells were pulsed with 3 μM puromycin for 1 h at 24 h posttransfection and then subjected to Western blot analysis or an IFA. Original magnification 200× (scale bars 100 μm). (C) EGFP was linked to the HA tag and E66L(13–34), and the resulting construct was named EGFP-E66L(13–34). (D and E) HEK-293T and IPI-2I cells were cotransfected with pRL-SV40 encoding the Rluc reporter gene downstream of the SV40 promoter and one of the following plasmids: PCAGGS, EGFP-HA, or EGFP-E66L(13–34)-HA. At 24 h posttransfection, the cells were lysed and subjected to Western blot analysis. The error bars show the SD of the results from three independent experiments; **, P < 0.01; ***, P < 0.001.