Fig. 9.
Ank9 and O. tsutsugamushi each inhibit host cell protein secretion. (A) Ank9 inhibits host cell protein secretion in a GLD-dependent and F-box independent manner. Culture media from HeLa cells co-expressing FLAG-tagged BAP, Ank9, Ank947–422 (lacks GLD), or AnkΔF-box together with luciferase was assayed for the presence of secreted luciferase. Each condition was transfected in quadruplicate and each cell supernatant assayed in quadruplicate. (B) O. tsutsugamushi impairs host cell protein secretion. HeLa cells transfected to express a secreted luciferase reporter were mock infected or infected with O. tsutsugamushi at an MOI of 5, 17.5, or 35. Cell culture media was collected and analyzed for secreted luciferase activity. Relative luciferase activity in the media recovered from infected cells was normalized to that in media from mock infected cells. Data are representative of three experiments with similar results. Statistically significant (**P < 0.01; ***P < 0.001) values are indicated. ns, not significant. Data is representative of three independent experiments with similar results.