Fig. 6. Immunofluorescence detection of RANTES binding to HMEC-1 cells. Scanning laser confocal microscopy was used to analyse the distribution of RANTES on the surface of HMEC-1 cells that had been cultured using a range of experimental conditions. (A) RANTES expression on the surface of resting HMEC-1 which had been treated with exogenous RANTES. (B) RANTES expression on the surface of HMEC-1 that had been stimulated with TNF- and IFN- and then treated with exogenous RANTES. (C) Expression of endogenous RANTES on the surface of HMEC-1 that had been stimulated with TNF- and IFN-. (D) RANTES expression on the surface of HMEC-1 cells that had been stimulated with TNF- and IFN- in the presence of chlorate and then treated with exogenous RANTES. (E) A constructed X-Z section through Fig. 6B confirming that binding of the exogenous RANTES occurs only on the apical surface of the cultured endothelial cells. (F) Summary data showing inverted mean fluorescence pixel intensity per cell for each of the four treatment groups; the resting and (cytokine) activated groups were treated with exogenous RANTES, the endogenous control group was cytokine-activated but received no exogenous RANTES and the chlorate control group was cytokine activated and received exogenous RANTES. The error bars show the s.e.m.