Figure 1

ADAM17-HA (AD17-HA) has mature form in SCC-9 cells and increases shedding of HB-EGF in HEK293 cells. A: SCC-9 cells were harvested, lysed with SDS sample buffer and immunoblotting was performed using anti-HA antibody. Anti-GAPDH antibody was used as loading control. B: SCC-9 cells were harvested and lysed in the presence and absence of BB-2516 and 1,10-phenanthroline. Immunoblotting was performed using anti-HA antibody and anti-GAPDH antibody was used as loading control. C: HEK293 cells overexpressing ADAM17-HA have increased shedding activity. HEK293 cells stably expressing a chimerical construct of HB-EGF fused with alkaline phosphatase (AP) were transfected with pcDNA-ADAM17-HA and released AP was quantified in cell supernatants after PMA (50 ng/ml) treatment. Two independent experiments with 3 replicates were performed (n = 2, Student’s t-test, DMSO: p = 0.0038, PMA: p = 0.0066). D: Immunoblotting of HEK293 cells overexpressing ADAM17-HA, performed using anti-HA antibody and anti-GAPDH antibody was used as loading control.