Figure 2. ADAMTS13 degradation by... | American Society of Hematology

Figure 2.

ADAMTS13 degradation by activated neutrophils. (A) ADAMTS13 was incubated with 0 × 103 to 500 × 103 PMA-stimulated neutrophils for 1 hour at 37°C, in the absence or presence of 2 mM AEBSF or 5 mM EDTA (B). (C) ADAMTS13 was incubated with 5 × 103 PMA-stimulated neutrophils with increasing concentrations of AEBSF, cathepsinG inhibitor I (CGI I), or Sivelestat for 1 hour. Samples were separated via SDS-PAGE and analyzed by (A) western blot using an anti-ADAMTS13 metalloprotease domain antibody, or (B-C) SYPRO Ruby. Black arrows indicate the level of full-length ADAMTS13 (∼180 kDa). AEBSF, 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride; EDTA, ethylenediaminetetraacetic acid.

ADAMTS13 degradation by activated neutrophils. (A) ADAMTS13 was incubated with 0 × 10³ to 500 × 10³ PMA-stimulated neutrophils for 1 hour at 37°C, in the absence or presence of 2 mM AEBSF or 5 mM EDTA (B). (C) ADAMTS13 was incubated with 5 × 10³ PMA-stimulated neutrophils with increasing concentrations of AEBSF, cathepsinG inhibitor I (CGI I), or Sivelestat for 1 hour. Samples were separated via SDS-PAGE and analyzed by (A) western blot using an anti-ADAMTS13 metalloprotease domain antibody, or (B-C) SYPRO Ruby. Black arrows indicate the level of full-length ADAMTS13 (∼180 kDa). AEBSF, 4-(2-aminoethyl)benzenesulfonyl fluoride hydrochloride; EDTA, ethylenediaminetetraacetic acid.

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