Figure 7.

(A) Production of CXCL-1 (KC) following infection (MOI = 10) in BHK cells with control TMEV (TV), control HEL-TMEV (HTV), and KC-TMEV (KCTV) analyzed 24-h post infection with Western blotting. Cell lysates in Tris-buffered saline with 1% Triton X-100 were separated by electrophoresis, transferred to nitrocellulose membranes, and then analyzed using rabbit anti-KC antibody. A representative result from two similar experiments is shown. (B) BHK cell lysates from triplicate cultures that were mock-infected, or infected with control TMEV or KC-TMEV for 6 and 24 h were subjected to a mouse KC-specific ELISA (BD, CA). The means (± SD) of the results from triplicate cultures are shown. (C) To further determine whether the KC produced after infection with KC-TMEV remained cell bound or was secreted, culture supernatants and cell lysates of BHK cells mock-infected or infected with KC-TMEV were assessed for the presence of mouse KC using ELISA. The means (± SD) of the results from triplicate cultures are shown. (D) Supernatants of monolayer BHK cells mock-infected or infected with control TMEV or KC-TMEV for 24 h were overlaid with Boyden chamber membranes containing GFP-expressing peripheral blood neutrophils that were enriched by using negative magnetic sorting. The cultures were incubated for 3 h and the membranes were flipped over to assess the number of transmigrated cells. An example microscopy image of an experiment is shown. A summary of three transmigration assessments is shown. Values given are the means (± SD) of the results from three separate experiments. ^***p < 0.001.