Altered lipid raft–associated signaling and ganglioside expression in T lymphocytes from patients with systemic lupus erythematosus
J. Clin. Invest. Elizabeth C. Jury, et al. 113:1176 doi:10.1172/JCI20345 [
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Figure 7The effect of T cell activation on GM1 expression and location of CD45. T cells from patients with SLE and healthy controls (Normal) were activated using magnetic beads coated with anti-CD3 and anti-CD28 for 1, 5, 10, 15, 30, and 60 minutes. Activated cells were subsequently fixed for confocal microscopy or lysed for analysis by Western blotting. (A) Figure representative of three experiments showing T cell expression of CTB during TCR activation. (B) T cells from patients with SLE or normal controls were allowed to “rest” overnight in complete medium, then were fixed for confocal microscopy analysis or were activated using magnetic beads coated with anti-CD3 and anti-CD28 for 5 minutes. Activated cells were then fixed for confocal microscopy. The images are representative of four experiments. The bar graph shows semiquantitation of CTB binding (MFI ± SEM) in five representative images examined from each of four patients with SLE and four healthy controls. (C) Figure representative of three experiments showing T cell expression of CD45 during TCR activation. (D) CD45/CTB overlay images comparing SLE and normal T cells after TCR activation; yellow color indicates areas of colocalization. (E) Proteins from T cell lysates were separated by 8% SDS-PAGE, transferred to PVDF membranes, and probed with antibodies against phosphotyrosine (pY) and active LCK (Src pY414) and against actin as a control for equal protein loading (Actin). Scale bars, 5 μm. X on confocal images represents position of activating bead.
