Figure 4.

Treatment with Dot1L inhibitor decreases nuclear translocation of NF-κB. (A) A549 cells were plated alone or with Dot1L inhibitor (1 μM); 48 h later, TNFα was added (10 ng/ml, 30 min). Cells were processed for immunofluorescence using anti-lamin and -p65 antibodies. (B) Cells were PR8hv-infected (m.o.i. 3, 8 h) with or without Dot1L inhibitor as in part (A). The inhibitor was present throughout the experiment in treated cells. The cells were processed for immunofluorescence using anti-NP, -lamin and -p65 antibodies. Nuclear NF-κB translocation was measured by orthogonal projection image analysis and quantified in at least 200 cells/condition. The ratio was calculated of p65 relative intensity in the nucleus and cytoplasm of each cell in different conditions. Ratios are shown in dispersion graphs beneath representative images for each experiment. Three independent experiments were performed.