Figure 5.

Effect of ADAMTS7 Genotype on ADAMTS7 Prodomain Processing

Cultured HEK293 cells were transfected with a plasmid to produce a recombinant protein containing the ADAMTS7 prodomain and metalloproteinase domain followed by a c-Myc epitope tag, with either serine (Ser) or proline (Pro) at residue 214 in the ADAMTS7 prodomain. Conditioned media, cell surface washes (with 0.5M NaCl), and cell lysates were subjected to immunoblot analyses with an anti-cMyc antibody or an anti-ADAMTS7 prodomain antibody. The same amount (40 μg) of proteins for each allele was loaded.

(A) A schematic diagram for the produced recombinant protein. P1, Pint, and M indicate the previously reported initially processed form, intermediately processing form, and fully processed form, respectively;⁴ CP indicates a cleaved ADAMTS7 prodomain fragment.

(B to G) Representative images of the immunoblot (IB) analyses and column chart presentations of mean (±SEM) values (n = 4 for each allele). ^∗p < 0.05 comparing ADAMTS7-214Ser and ADAMTS7-214Pro.