Figure 7: ADAMTS17-PCD does not cleave fibrillin-1 or fibrillin-2. | Scientific Reports

Figure 7: ADAMTS17-PCD does not cleave fibrillin-1 or fibrillin-2.

From: Unusual life cycle and impact on microfibril assembly of ADAMTS17, a secreted metalloprotease mutated in genetic eye disease

Figure 7

(a) Coomassie-blue stained SDS-PAGE of fibrillin-1 (FBN1-N, FBN1-C) and fibrillin-2 (FBN2-N, FBN2-C) peptides (5 μg/lane) (see Fig. 6a) incubated with (+) or without (−) recombinant ADAMTS17-PCD (5 μg/lane) shows no evidence for proteolysis of fibrillin-1 or fibrillin-2 in vitro. M = mature enzyme; Z = zymogen. (b) Western blot analysis of conditioned medium (Med) and cell lysate (Lys) from HEK293F cells stably expressing fibrillin-1 and transiently expressing ADAMTS17 (17), ADAMTS17EA (17-EA), ADAMTS17-PCD (PCD) or empty vector (v). Western blots were probed with antibodies against fibrillin-1 (anti-FBN1-C, green) and the recombinant ADAMTS17 peptides (anti-myc, red). (c) Western blot analysis of conditioned medium (Med) and cell lysate (Lys) from HEK293F cells stably expressing FBN2-N or FBN2-C with ADAMTS17 (17), ADAMTS17EA (17-EA), ADAMTS17-PCD (PCD), or empty vector (v) shows no evidence of cleavage. Western blots were probed with antibodies detecting fibrillin-2 (anti-FBN2-N or anti-FBN2-C, green) and the recombinant ADAMTS17 peptides (anti-myc, red).

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