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. 1998 Nov 16;188(10):1795–1802. doi: 10.1084/jem.188.10.1795

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Antagonism of Fas-induced apoptosis in normal thymocytes by Con A activation was mediated by MKK. (A) Freshly isolated thymocytes from 8-wk-old MRL +/+ and MRL lpr/lpr mice were stimulated with immobilized anti-Fas antibody Jo2 (coated at 5 μg/ml) and/or Con A (4 μg/ml) in the absence or presence of PD 098059 (PD, 10 μM) for 4 h. Cell death was determined by Annexin V (Clontech, Palo Alto, CA) staining. Data shown are thymocytes from +/+ mice except Jo2 (lpr). The result is the average of duplicates ± SD. Experiments were repeated twice with identical results. (B) Induction of FLIP was MKK-dependent in thymocytes. +/+ thymocytes were activated with Con A in the absence or presence of 10 μM PD 098059, and total RNA was isolated 1, 2, and 3 h after activation. FLIP transcript was determined as described in the legend to Fig. 3 except primers for mFLIP were used during PCR analysis.

Antagonism of Fas-induced apoptosis in normal thymocytes by Con A activation was mediated by MKK. (A) Freshly isolated thymocytes from 8-wk-old MRL +/+ and MRL lpr/lpr mice were stimulated with immobilized anti-Fas antibody Jo2 (coated at 5 μg/ml) and/or Con A (4 μg/ml) in the absence or presence of PD 098059 (PD, 10 μM) for 4 h. Cell death was determined by Annexin V (Clontech, Palo Alto, CA) staining. Data shown are thymocytes from +/+ mice except Jo2 (lpr). The result is the average of duplicates ± SD. Experiments were repeated twice with identical results. (B) Induction of FLIP was MKK-dependent in thymocytes. +/+ thymocytes were activated with Con A in the absence or presence of 10 μM PD 098059, and total RNA was isolated 1, 2, and 3 h after activation. FLIP transcript was determined as described in the legend to Fig. 3 except primers for mFLIP were used during PCR analysis.