(a) Effects of E-64 in fibroblasts. CD55−/−cath-D and CD55−/−D231Ncath-D cells were either untreated or treated for 48 h with 10 mM E-64 and cellular cath-D was analysed by immunoblotting.
(b) Concentration dependency of the E-64 effects. CD55−/−cath-D cells were either untreated or treated for 48 h with increasing concentrations of E-64 and cellular cath-D was analysed by immunoblotting.
(c) Effects of E-64 using two-dimensional electrophoresis. CD55−/− cath-D cells were either untreated or treated for 48 h with 10 mM E-64 and cellular cath-D was analysed by two-dimensional electrophoresis and immunoblotting.
(d) Effects of CA-074Met. CD55−/−cath-D and CD55−/−D231Ncath-D cells were untreated or treated for 48 h with 50 μM CA-074Met and cellular cath-D was analysed by immunoblotting. The effect of 10 mM E-64 is shown.
(e) Effects of CLICK-148 alone or in combination with CA-074Met. CD55−/−cath-D and CD55−/−D231Ncath-D cells were either untreated or treated for 48 h with 5 or 20 μM CLICK-148 alone or in combination with 50 μM CA-074Met. Cellular cath-D was analysed by immunoblotting.
Brackets indicate the 52-48 kDa processing intermediates detected in CD55−/−cath-D cells and the 51-47 kDa processing intermediates detected in CD55−/−D231Ncath-D cells.