Inhibitory role of TACE/ADAM17 cytotail in protein ectodomain shedding

Figure 2 L-selectin ectodomain shedding activity in cytotail-deleted tumor necrosis factor-α converting enzyme constructs with cysteine to alanine substitutions. The shedding-defective M1-L-selectin cells, which stably express transmembrane L-selectin, were transiently cotransfected with the transfection marker green fluorescence protein (GFP), and indicated tumor necrosis factor-α converting enzyme (TACE) vectors or the control pRK5 plasmid at a ratio of 1:3. Transmembrane L-selectin on the surface of live unfixed cells was immunostained with anti-L-selectin and a PE-conjugated secondary antibody, and was detected by flow cytometry. GFP-positive cells were gated as cells expressing the cotransfected TACE construct. Green: ∆C TACE; Red: ∆C TACE with a cysteine to alanine substitution at indicated position; dotted blue lines, pRK5.