FIG. 1.

Analysis of N1L expression and viral growth. (A) Synthesis of N1 protein was assessed in BS-C-1 cells infected with ECTV wt (wt) or one of three clonal isolates of ECTVΔN1L (ΔN1L #1 to #3) or ECTV-N1Lrev (N1Lrev #1 to #3) at 24 h p.i. (MOI, 5). Mock-infected cells (mock) served as a control. Cell lysates were prepared and separated in a 12% SDS-polyacrylamide gel. After the proteins were transferred to nitrocellulose membranes, the N1 protein and β-actin were detected using an N1-specific polyclonal rabbit antiserum and β-actin-specific monoclonal mouse antibody, respectively. (B and C) Multiple-step growth analysis of ECTV wt, ECTVΔN1L, or ECTV-N1Lrev was performed with BS-C-1 (B) and NIH 3T3 (C) cells infected at an MOI of 0.01. Data shown are mean values from two or three independent experiments.