Orientia tsutsugamushi uses two Ank effectors to modulate NF-κB p65 nuclear transport and inhibit NF-κB transcriptional activation
Fig 5
O. tsutsugamushi inhibits NF-κB-dependent gene transcription.
Stably transfected HeLa cells bearing four copies of the NF-κB response element upstream of the Firefly Luciferase (Luc) gene seeded in a 96-well plate were incubated with O. tsutsugamushi at an MOI of 10, 50, or 100 or mock control for 24 h. The cells were treated with TNFα or vehicle control for 8 h to promote NF-κB-dependent gene transcription followed by the addition of lysis buffer and Luc substrate. The plate was read in a luminometer for 10 s per well to quantify Luc activity. Data are presented as the mean percentage + SD of Luc activity normalized to the Luc activity of mock control cells exposed to TNFα from triplicate samples. Statistically significant (***P < 0.001; ****P < 0.0001) values are indicated. Data are representative of three independent experiments.