Figure 2

Western blotting on U2-OS cell lysates. A. Lane 1 and 6: marker. Lane 2, 4 and 7: U2-OS cell lysates; Lane 3, 5 and 8: RIPA buffer. 5 bands have been identified in U2-OS cell lysates by NB509-11G8. Moreover, these bands can be completely blocked by the selection peptide, but not the truncated peptide. B. U2-OS cell lysates in vitro digestion products detected by X53 on western blotting. Lane1: marker. Lane 2: U2-OS cell lysates; Lane 3–6: U2-OS cell lysates incubated with 5 ug/ml collagenase for 1 hr, 2 hr, 4 hr or overnight. Lane7-10: U2-OS cell lysates incubated with 50 ug/ml collagenase for 1 hr, 2 hr, 4 hr or overnight. X53 only detected 64 kDa α1 chain. The intensity of this chain decreased with the incubation time and the amount of collagenase. C. U2-OS cell lysates in vitro digestion products detected by NB509-11G8 on western blotting. Lane 1: U2-OS cell lysates; Lane 2–5: U2-OS cell lysates incubated with 5 ug/ml collagenase for 1 hr, 2 hr, 4 hr and overnight. Lane6-9: U2-OS cell lysates incubated with 50 ug/ml collagenase for 1 hr, 2 hr, 4 hr or overnight. Lane10: marker. The degradation of ColX by collagenase exhibited a time- and dose- dependent pattern.