Fig. 3.

Tat induced increase in CXCL10 from R28 cells lead to increased migration of monocytes. Analysis of chemotactic activity of CXCL10 in supernatants collected from R28 cells treated with Tat, in absence and presence of AMG487, blocker of CXCL-10 receptor on monocytes, showed pronounced contribution of CXCL10. IFN-γ was used as positive control. Supernatant from unstimulated control cells and from IFN-γ and Tat-treated R28 cells were added to the lower chamber of ChemoTX microplate. 2×10⁶ PBMCs were placed on the top of the membrane. In indicated wells, PBMCs were treated with two different concentrations of CXCR3 blocker, AMG487 for 2 h before loading in the top chamber of the ChemoTX microplate. After 4 h, PBMCs migrating towards supernatant collected from different treatments in lower chamber were counted by using Cell Titer 96 Aqueous One Solution Assay. Migratory response of PBMCs to supernatant fluids are shown as migration index, calculated as the number of cells migrating towards the treated supernatants divided by the number of cells migrating toward the medium only