a) Predicted docked complex of Rv2615c (yellow) bound to TLR2 (green) and TLR4 (blue) using HADDOCK server. b) THP-1 macrophages were stimulated with control/Rv2615c protein for 24h and 48h. Expression profile of TLR2, TLR4 and HLA-DR was evaluated at 24h and 48h of stimulation. The percentage of TLR/HLA-DR positive cells was plotted on the y axis and control/Rv2615c protein on the x axis of graphs for various time points. The findings of the statistical study, which used the Student’s t test, are shown as Mean±SEM for three separate experiments where * represents comparison with unstimulated cells and *P<0.05, **P<0.01, ***P<0.001. c) THP-1 cells were blocked with Anti-TLR4 antibody prior to stimulation and evaluated for levels of TLR4 and HLA-DR at 24h. Isolated mRNA from unstimulated or Rv2615c/LPS stimulated macrophages were synthesized into cDNA and RT-PCR was performed to estimate the levels of d) MyD88 and e) NF-κB gene expression using 2-∆∆ CT method. MyD88 and NF-κB gene expression in unstimulated cells was considered 1 and with respect to unstimulated cells fold change in MyD88 and NF-κB gene expression in LPS and Rv2615c was plotted. Graphs were plotted for different time points with fold change in MyD88/NF-κB expression on y axis and LPS/Rv2615c protein on x axis. Results are expressed as Mean ± SEM of three independent experiments.