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. 2008 Jun 16;28(16):5014–5026. doi: 10.1128/MCB.00640-08

FIG. 2.

FIG. 2.

MHC-II signaling of cell death does not require activation of Src family tyrosine kinases. (A) K46 cells were treated with 25 μM PP2 or dimethylsulfoxide (DMSO) for 30 min at 37°C and then stimulated with biotin-MHC-II (M5.114; 20 μg/ml) and avidin (20 μg/ml) for the indicated times. SDS-PAGE-fractionated whole-cell lysates were probed with the indicated Abs (n = 3). (B) The intracellular free calcium concentration ([Ca2+]i) was measured before and during K46 stimulation with biotin-MHC-II (M5.114; 10 μg/ml) and avidin (10 μg/ml). Cells were pretreated with DMSO or PP2 as described for panel (A) (n > 3). (C) K46 cells treated as for panel A were stimulated with biotin-anti-MHC-II (M5.114; 10 μg/ml; shaded bars) or biotin-anti-FcγRIIB (2.4G2; 10 μg/ml; open bars) and avidin (10 μg/ml) for 5 h, and death was measured by annexin V-PI staining as described in Materials and Methods (n > 3). Error bars represent standard deviations of triplicates.