Fig 5.

Verification of NL63 replication in PESU-B5L cells. Total RNA was harvested from cells on day 5 postinfection and subjected to RT-PCR using primers designed to detect leader-containing transcripts of the N gene, which would be present only if NL63 replicated. (A) Subgenomic transcripts were detected in the PESU-B5L cells infected with both wtNL63 and NL63gfp. (B) The sgN band for wtNL63 was excised, and the DNA was purified and sequenced to verify that the HCoV-NL63 leader sequence and the 5′ end of the N gene were present. L, ladder; N, wt-NL63; G, NL63gfp; M, mock. The leader sequence has a beige bar beneath it, and the 5′ end of the N gene has a green bar beneath it.