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Fig. 1. Infection with T. gondii fails to induce NF-
B-dependent gene expression and does not result in nuclear accumulation of NF-B. (A) Clonal 3T3 cells expressing GFP driven by a promoter containing NF-B binding sites were used to assess NF-B responsiveness during T. gondii infection. Cells were either left untreated (left panel), stimulated with TNF-
(10 ng/ml) for 18 hours (middle panel), or infected with p30-RFP-expressing Rh parasites for 18 hours (right panel); parasites were generated as previously described (Striepen et al., 2001). Expression of GFP and RFP was assessed by flow cytometry using BD FACS Calibur and analyzed using Cell Quest (Becton Dickinson, Mountain View, CA). (B) Indirect immunofluorescence analysis of T. gondii-infected HFF cells demonstrates lack of nuclear accumulation of NF-B. Anti-p65 (red) staining reveals cytoplasmic localization in uninfected cells and nuclear accumulation in cells stimulated with TNF- for 1 hour. `DAPI' indicates staining of host cell and parasite nuclei (blue). Yellow triangles indicate parasite-containing vacuoles, and infection times are indicated in the top left corner of each panel. The absence of nuclear accumulation of NF-B in infected cells correlates with the lack of GFP expression in panel A.
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