Figure 2.

Chemical cross-linking of IN1-105. The reaction was carried out in 50 mM Hepes pH 7.8. IN 1 –105 and BS³ was incubated at 25°C in 40 μL at the times and concentrations indicated. The reaction was stopped with 50 mM lysine pH 8.0. (A) Crosslinking kinetics. 56 μM of IN 1–105 were incubated with 100 μM of BS³, aliquots were removed at the times indicated. Left, SDS-PAGE (15% acrylamide) silver stained (4 μg of protein were loaded in each lane). Right, formation of crosslinked IN 1–105 dimer (percent); quantification of dimeric and monomeric band were carried out with the imageJ program. (B) Effect of BS³ in crosslinking. 56 μM IN 1–105 and BS³ (0 to 300 μM) were incubated for 35 min at 25°C, aliquots containing 4 μg were loaded on a 12% acrylamide gel. Left, SDS-PAGE (12%) Coomasie brilliant blue stained gel. Right, formation of crosslinked IN 1–105 dimer.