For IFNγR binding experiments, a flow cell of a CM4 sensor chip was EDC/NHS activated as described above, after which 50 µL of Streptavidin (Sigma-Aldrich) at 200 µg/mL in 10 mM sodium acetate buffer, pH 4.1, were injected.
CD4+ T cells were isolated and activated as described above.
Human Umbilical Vein Endothelial Cells grown on coverslips were starved and activated as described above.
The data used in this study (read counts, library sizes, and estimated dispersion parameters) are given in Additional file 1. Naive T cells were isolated and activated as described above.
A total of 5×10 BMMΦ were activated as described above in a final volume of 1 mL; 18 h later, 25×10 L. major parasite were added to the cultures and 2 days later, the plates were washed with PBS to remove non-phagocytosed promastigotes and the MΦ were lysed as described in 45.
The signal from a blank flowcell, (which had been activated and blocked as above, but to which no peptide had been immobilized) was subtracted from all SPR measurements to correct for bulk solvent effects.
Most genes advanced by LasR may be activated indirectly as mentioned above, or LasR binding sites may be more complex.
CD4+ T cells were activated overnight as described above, collected, washed, and transferred to new wells and cultured for another 48 hrs.
aCD4 that were activated overnight as described above were collected, washed, and cultured in 24-well plate at 1 × 10, 2 × 10, or 4 × 105 cells/2 ml of CM for 48 hr to generate conditioned media (hereafter called 5 × 10, 1 × 105 and 2 × 105 aCD4S, respectively).
