Direct alteration of NFκB activity near the time of infection is sufficient to modulate the frequency of non-productive RGH infection. A. Experimental outline for doxycycline-inducible NFκB modulation and RGH infection of Jurkat cells. B: Jurkat cells bearing a stably integrated, doxycycline-inducible expression construct containing either a dominant negative IκBα or p65 open reading frame were infected with RGH. Cells were cultured with or without doxycycline for 24 hours, and then washed and cultured in fresh media until analysis by flow cytometry at four and seven days post-infection. Error bars represent standard deviations of triplicate experiments. ‘ns’ non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001 (Student’s T test). C: Immunoblot analysis of cells used in panel B. Jurkat whole cell extracts were separated by SDS-PAGE, transferred to nitrocellulose membrane, and blotted with antibodies against IκBα or p65.