Figure 6. The effect of infection of HEK293T, 293/TLR2, and 293/TLR4/MD-2 cells with live R. akari on activation of IRAK1 and p38.

HEK293T cells and 293/TLR2 and 293/TLR4/MD-2 stable transfectants were incubated for 2 h with medium or infected with R. akari at MOI = 50. (A) Cell lysates were prepared and subjected to immunoblot analyses with Ab against p-IRAK1 and p-p38 to analyze activation of these kinases based on their phosphorylation, and anti-tubulin Ab was used to control for protein loading. (B) Bacterial loads were estimated based on PCR analysis of 16S R. akari RNA in samples of infected cells. No RT indicates PCR reaction on the RNA sample isolated from R. akari-infected HEK293T cells in the absence of RT. Shown are data of a representative experiment.