Distribution of IP-10 and RANTES. (A) HUVECs were activated by TNF and IFN-γ before fixation and paired immunostaining for RANTES (clone ID2/A12) and VWF (top panels) or IP-10 (clone 33036.211) and VWF (bottom panels). Original magnification, × 100. Corner insets show high magnification of framed areas. (B) Cell-associated levels of RANTES and IP-10 in TNF- and IFN-γ-activated HUVECs were measured by ELISA after incubation without or with PMA (100 ng/mL, 1 hour). The response of IL-8 toward PMA was examined in parallel wells. Secretory ratios were calculated as explained in the Figure 1 legend, with the dashed line indicating a ratio of 1 and absence of regulated secretion. (C) TNF- and IFN-γ-activated HUVECs incubated without (-) or with (+) PMA (100 ng/mL, 1 hour) before fixation and paired immunostaining for RANTES (BAF278; left panels) and VWF (corner insets) or for IP-10 (clone 33036.211; right panels) and VWF (not shown). Original magnification, × 40.