Figure 3.

Fodrin cleavage promotes CD45 clustering and membrane blebbing during galectin-1 T cell death. A) (left) Jurkat cells were treated with buffer control (C) or galectin-1 (G) for 60 min in the absence or presence of calpain inhibitor IV. CD45 (green) and fodrin (red) were detected by immunofluorescence confocal microscopy. (right) CD45 (green) and annexin V (A5) binding were detected on cells treated as above. CD45 clustering, fodrin redistribution, and membrane blebbing were reduced in the presence of calpain inhibitor IV, compared to cells treated with galectin-1 alone. Annexin V binding was evident on cells treated with calpain inhibitor IV, but clustering on blebs was reduced compared to cells treated with galectin-1 alone. B) Jurkat and J45.01 cells were treated with buffer control (C) or galectin-1 (G), and CD45 and fodrin (left), or CD45 and annexin V binding (right), detected as in (A). No CD45 staining was detected on J45.01 cells. Fodrin redistribution, membrane blebbing, and annexin V clustering were markedly reduced in J45.01 cells compared to Jurkat cells. Data are representative of three independent experiments.