Figure 2
GzmB Cleavage assays on dermal-epidermal junction proteins. (a) 4–20% SDS-PAGE western blot of GzmB-mediated cleavage of α6 integrin (α6 int) sub-unit with and without inhibitors serpin A3N (SA3N) and compound 20 (Com20). Black arrows indicate cleavage fragments and * indicates full-length proteins. At a concentration of 200 nM GzmB produces cleavage bands, and this cleavage is prevented by GzmB inhibitors. Full-length blot is presented in Supplemental Fig. S5. (b) 4–20% SDS-PAGE western blot of GzmB-mediated cleavage of β4 integrin sub-unit (β4 int) with and without inhibitors SA3N and Com20. Black arrows indicate cleavage fragments and * indicates full-length proteins. At a concentration of 200 nM GzmB produces cleavage bands, but its inhibition prevents the appearance of these bands. Full-length blot is presented in Supplemental Fig. S6. (c) 10% SDS-PAGE western blot of GzmB-mediated cleavage of collagen VII (coll VII) with and without inhibitors SA3N and Com20. Black arrows indicate cleavage fragments and * indicates full-length proteins. GzmB produces cleavage bands, and this cleavage is reduced by the addition of Com20 and abolished by SA3N. Full-length blot is presented in Supplemental Fig. S7. (d) 8% SDS-PAGE western blot of GzmB-mediated cleavage of collagen XVII (coll XVII) with and without inhibitors SA3N and Com20. Black arrow indicates cleavage fragment and * indicates full-length protein. GzmB produces a cleavage band, and this cleavage is abolished by the addition of Com20 or SA3N. Full-length blot is presented in Supplemental Fig. S8.
