Figure 8From: Association of CD99 short and long forms with MHC class I, MHC class II and tetraspanin CD81 and recruitment into immunological synapsesCD99 is recruited into the IS and lipid rafts. (A) DDAO-SE pre-labeled Jurkat T cells were incubated with SEB-loaded Raji B cells. Cells were then plated onto a cover slip, fixed, stained with anti-CD99 mAb followed by Alexa Fluor 488-conjugated goat anti-mouse IgG antibodies, TRITC-phalloidin for F-actin and Hoechst 33258 for the nucleus. The stained cells were visualized by confocal fluorescence microscopy. The green and yellow images represent the localization and distribution of CD99 and F-actin, respectively. The nuclei and Jurkat cells are shown in blue and red, respectively. (B) Jurkat cells were homogenized to isolate cell membranes, which were then solubilized in lipid raft preserving detergent 1% Brij-98 or lipid raft disrupting detergent 1% Lauryl maltoside (LM) and subjected to gel filtration on Sepharose 4B. The obtained fractions were analyzed by Western immunoblotting using anti-CD99 and anti-Lck mAbs.Back to article page