Figure 4

Type I strains differentially activate NF-kB nuclear translocation, and is dependent on GRA15. (A) HEK293T NF-kB reporter cells were infected with RH-ERP, RH-JSR, RH+GRA15_GT1 and GT1. For each strain, the luciferase reading after 24 hours infection was taken. The graph shows averages from three independent experiments, with levels representing fold change of NF-kB luciferase readings normalized to uninfected, unstimulated control cells, and the error bars represent the standard error. Mean + SEM, of three experiments, * p-value < 0.01, N.S. not significant, Student’s t-test. (B) HFFs were infected for 24 h with RH-ERP, RH-JSR, GT1 and RH+GRA15_GT1 transgenic, uninfected cells were stimulated with TNF-alpha for 1 h (TNF), or left unstimulated and uninfected (UI). Cells were fixed, probed with p65 antibody and mean nuclear staining was quantified, the error bars represent the standard deviation, * p-value < 0.05, Student’s t-test. Quantification shown is representative of three independent experiments. (C) IL12-p40 and (D) CCL2 levels were measured in supernatants from C57BL/6 bone marrow derived macrophages infected for 24 hours with RH-ERP, RH-JSR, RH+GRA15_GT1 transgenic and GT1. The graphs shown are representative of three independent experiments for IL-12p40 and two independent experiments for CCL2, and the error bars represent the standard deviation, * p-value < 0.01, Student’s t-test between strain and RH-ERP infected BMMs, + p-value < 0.01, Student’s t-test between the two conditions indicated.