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Fig. 7. ATP--S-induced HB-EGF shedding and EGFR activation are sensitive to ADAM inhibition. (A,B) Cells expressing HB-EGF-AP were pretreated with (A) 50 µM GM6001 (GM) or its inactive analog (GM neg) or (B) 4 µM GW280264X (GW) for 1 hour and stimulated with 100 µM ATP--S. AP activity was measured and expressed as in Fig. 5. ^**P<0.01, significant decrease in AP release by the inhibitors in the control and ATP--S-treated cells. NT, no inhibitor treatment. (C) Growth-factor-starved THCE cells were pretreated with 10 µg/ml CRM197 (CRM), 4 µM GW280264X, 50 µM GM6001 or 50 µM GM6001 inactive analog for 1 hour and then stimulated with 100 µM ATP--S for 10 minutes. Cells were then lysed and lysates were subjected to EGFR immunoprecipitation as described in Fig. 3A and phosphorylation determination.

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