Fig. 2. The T.-gondii-mediated translocation of NF-B occurs early in infection. MEFs were infected at a m.o.i. of 5:1 for 1 hours, 2 hours, 3 hours, 4 hours, 5 hours and 6 hours. Double immunofluorescence was performed with primary rabbit polyclonal antibodies against NF-B p50 (A) and p65 (B) and mouse monoclonal anti-T.-gondii SAG1. Translocation of p50 and p65 occurred as early as 1 hour p.i. (A,B, top, respectively). Scale bar, 20 µm. Single-cell analysis of NF-B translocation during the 6 hour incubation period showed significant differences in the proportions of infected cells displaying p50 and p65 nuclear staining compared with uninfected cells (A,B, bottom, respectively). Cells were counted in a minimum of 20 microscopic fields under 100x and numerical data was collected from three separate experiments. *, P<0.05.