FIG 6.

The GSK3/PI3K/mTOR pathway plays an important role in E. chaffeensis survival. (A) THP-1 cells were infected with E. chaffeensis (MOI of 50). Bacterial loads were determined at 3, 24, and 48 h posttreatment with pyrvinium by measuring the percentage of infected cells by counting 100 Diff-Quik-stained cells. Data represent means ± SD (n = 3) (P < 0.05). (B) THP-1 cells were transfected with specific or control siRNA to knock down Rheb/p70S6K/PTEN and were then infected with E. chaffeensis (1 day posttransfection). Ehrlichial infection was determined using qPCR measurement of the dsb copy numbers at 24 and 48 h pi. Data are represented as means ± SD (*, P < 0.05, n = 4). (C) Western immunoblot to confirm reduction of Rheb, p70 S6K, and PTEN protein levels after siRNA transfection.