LFA-1

1. A LFA-1 Rab11 Merge B 0 min 20 min Supplemental material S7. (A) Cytofluorimetric analysis of surface-expressed L/2 upon co-transfection with the Rab11S25N mutant. 1Cells were transfected with either GFP or the dominant negative GFP-Rab11S25N mutant and co-transfected with either L/2 or CD4. 24 hours post-transfection cells were surface-stained with anti-L or anti CD4 mAb followed by Cy3-conjugated secondary antibody. Double-fluorescent cells were gated and their red fluorescence values were analyzed. 2Mean Channel Fluorescence, expressed as Arbitrary Units. Data represent the mean ± S. D. of three separate experiments. 3p<0.01. (B) Rat Basophilic Leukemia (RBL) cells stably expressing the fMLP receptor were treated for 10 min on ice with an anti-rat CD18 mAb (WT-3), followed by stimulation with 100 nM fMLP and incubation for the indicated time points. Cells were then fixed, permeabilized and stained with a Cy3-conjugated anti-mouse IgG antiserum to detect beta-2 integrins and with a rabbit anti-Rab11 antiserum, followed by a FITC-conjugated goat anti-rabbit Ig antiserum. Nuclei were stained by Hoechst 33342.