FIG. 4.

WNV induces eIF2α phosphorylation and CHOP in primary rat hippocampal neurons. (A) One-week-old primary rat neurons were infected with WNV at an MOI of 10. At 24 h p.i., lysates were prepared and analyzed for phospho-eIF2α and total eIF2α by Western blotting. (B) Total RNA was isolated from WNV-infected neurons at 24 h p.i., and chop mRNA levels were quantitated by real-time RT-PCR. The chop message was normalized to the ribosomal protein L32 message, and x-fold changes were calculated as described in Materials and Methods. WNV infection was verified by WNV NS1 amplification from the samples. ND, not detected. Data represent three independent experiments and are means ± SD.