Figure 3

Time course study of SEB binding to RPTECs: Fluorescence-based co-reporting of SEB and CD1d: 1 x 10 ⁶ cells/ml RPTECs were exposed to 100 μg/ml SEB, washed and then CD1d and SEB were sequentially reported. To report CD1d, mouse anti-CD1d antibodies coupled to Alexa 594-conjugated goat anti-mouse antibodies were used (Red spots). To report SEB, rabbit anti-SEB antibodies coupled with FITC-conjugated goat anti-rabbit antibodies were used (Green spots). Control: The “no SEB” treatment was similar to Figure 2 (not shown herein); SEB (15 m): SEB assault for 15 minutes; SEB (30 m): SEB assault for 30 minutes; SEB (1 h): SEB assault for one hour; and SEB (2 h): SEB assault for two hours. All images are at 60X magnification imaged with a fluorescent microscope equipped with a digital camera (Olympus Optical Company).