Data analysis was performed using manufacturer’s program and is based on the ddCt method, with normalization of the raw data to the panel of housekeeping genes provided in the array. The genes showing modulation find more by 1.5 fold up or down were only www.selleckchem.com/products/U0126.html selected for further analysis. Functional annotations of the selected genes were carried out by the
bioinformatics software David for Bioinformatics. Three independent experiments with a pool of 2 donors each were analyzed. Statistical analysis Statistical evaluation of the data was done using GraphPad Prism 5 software. Student t-test was performed for simple comparison between 2 means. For multiple comparisons, the results were analysed by two-way ANOVA followed by Bonferoni’s post-test. p < 0.05 was considered statistically significant. All shown data are representative for at
least 3 independent experiments. Results Chlamydia trachomatis infect monocytes and monocyte-derived DCs in a comparable manner Monocytes isolated from human peripheral blood mononuclear cells (PBMCs) and monocyte-derived DCs were infected with C. trachomatis serovars Ba, D and L2 (Figure 1). Results show that all the three serovars were capable of infecting both the monocytes and DCs and form Tariquidar clinical trial inclusions as detected by immunofluorescence microscopy 2 days post infection (p.i.). However, the inclusions were smaller in size compared to typical inclusions that have been reported in
HeLa cells (Additional file 2: Figure S2). The inclusion morphology and staining intensity varied between the infected monocytes and DCs. Figure 1 Immunofluorescence microscopy of infected monocytes and monocyte-derived Dendritic cells (DCs). Monocytes (upper panel) and monocyte-derived DCs (lower panel) were infected with C. trachomatis serovars Ba, D and Clostridium perfringens alpha toxin L2 (MOI-3) for 2 days. Chlamydial inclusions (green) were stained with FITC conjugated anti-chlamydia LPS antibody and counterstained with Evans Blue. Pictures were taken at 63X magnification with Leica DMLB. The figures are representative of 3 independent experiments. In monocytes, the percentage of infected cells were comparable among the three serovars and did not seem to change even when the infection duration was extended to 3 days (Table 1). For DCs, the percentage of infected cells were similar for serovars Ba and D but serovar L2 showed a higher infection rate as compared to the other two (Table 1). However, the infection rate declined remarkably for all the three serovars when infected for 3 days. The infection rate was nevertheless much lower in both monocytes and DCs than in HeLa. Mock controls were prepared for each round of experiments which showed absence of chlamydial antigens in the donors (Additional file 3: Figure S3). Table 1 Comparison of infection rate in monocytes and monocyte-derived DCs infected with C.