Figure 4

U83A treatment results in long-term delays of CCL4 driven internalisation of CCR5 in the absence of re-stimulation. CCR5 expression in MAGI-CCR5 cells was monitored by flow cytometry after incubation with 100 nM U83A in buffer (plain line) or control, buffer only (dashed line) for 10 minutes, then washed, incubated for the indicated time (x axis, in minutes), and stimulated with 100 nM CCL4 for 10 minutes, cells were then stained with FITC-CCR5 antibody and fixed. CCR5 surface levels were then determined by flow cytometry. Results were normalised in reference to CCR5 level of expression at time 0, as 100% values, and are a combination of two experiments each run in duplicate. P values for mean differences are 0.029, 0.066 and 0.016 for 30, 60 and 120 minutes time points, respectively (unpaired T test).