Fig. 8.
Fig. 8. Discrete regions in GM-CSF and βc mediate their direct interaction. / Purified sβc was incubated at 25°C for 1 hour alone or in the presence of either GM-CSF or E21R. Samples were left untreated (A) or were treated for 10 minutes with BS3 cross-linker (B). To determine if GM-CSF was interacting with the cytokine-binding site in the fourth domain of βc, purified sβc was preincubated with a Fab fragment of the neutralizing anti-βc mAb, BION-1, or the nonneutralizing control anti-βc mAb, 2H1. GM-CSF was then allowed to bind and the mixture and individual mAb treated with BS3cross-linker (C). Samples were analyzed on 12.5% (A) or 10% (B-C) SDS-PAGE gels and stained with Coomassie. The positions of molecular weight markers are shown in kilodaltons, and the position of sβc cross-linked to GM-CSF is indicated by ◂.

Discrete regions in GM-CSF and βc mediate their direct interaction.

Purified sβc was incubated at 25°C for 1 hour alone or in the presence of either GM-CSF or E21R. Samples were left untreated (A) or were treated for 10 minutes with BS3 cross-linker (B). To determine if GM-CSF was interacting with the cytokine-binding site in the fourth domain of βc, purified sβc was preincubated with a Fab fragment of the neutralizing anti-βc mAb, BION-1, or the nonneutralizing control anti-βc mAb, 2H1. GM-CSF was then allowed to bind and the mixture and individual mAb treated with BS3cross-linker (C). Samples were analyzed on 12.5% (A) or 10% (B-C) SDS-PAGE gels and stained with Coomassie. The positions of molecular weight markers are shown in kilodaltons, and the position of sβc cross-linked to GM-CSF is indicated by ◂.

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