Fig. 3.

InlC prevents the translocation of NF-κB-p65 to the nucleus in infected cells. (A) HeLa cells noninfected (NI) or infected by Listeria monocytogenes WT, the ΔinlC mutant (ΔinlC), or the complemented-ΔinlC mutant (ΔinlC + inlC) were stimulated with TNF-α (50 ng/mL) for 30 min, fixed, and labeled by immunofluorescence with DAPI for the nucleus and bacteria, anti-InlC antibodies to detect the secretion of InlC in the cytoplasm of cells, and anti-p65 to recognize the NF-κB subunit. (B) To quantify the results shown in A, the percentage of nuclear translocation of NF-κB (P65) was determined by counting at least 100 noninfected or WT, ΔinlC, or ΔinlC+inlC infected cells using fluorescence microscopy. The experiment was performed twice, and typical results are shown. ***P < 0.0005, χ² test.