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. 2003 Feb 1;31(3):911–921. doi: 10.1093/nar/gkg186

Figure 7.

Figure 7

Interaction of recombinant EGR-1 and EGR-4 proteins with NFATc. (A) Recombinant GST-NFATc protein expressed in E.coli was coupled to GST. Various wash and elute fractions were obtained, separated by SDS–PAGE, transferred to nitrocellulose and assayed by western blotting using the indicated antibodies. The individual lanes represent the recombinant protein (lane 1), the material obtained after loading the column (lane 2), the flow through, (lane 3), several wash (lanes 4, 5 and 6) and the elute fraction (lane 7). (B) Interaction of EGR-1 and NFATc. EGR-1 expressed in insect cells (lane 1) was applied to an NFATc matrix and wash and elute fractions were assayed using an EGR-1 specific antiserum. (C) Interaction of EGR-4 and NFATc. EGR-4 expressed in insect cells (lane 1) was added to the NFATc matrix and wash and elute fractions were assayed with an EGR-4 specific antiserum. (D) As control an unrelated recombinant protein (SCRs 15–20 of complement factor H) (lane 1) was assayed for binding and the individual fractions were assayed using an anti factor H antiserum. (E) To exclude non-specific binding of EGR-1 and (F) of EGR-4, the GST matrixes were incubated with the recombinant EGR proteins and wash and elute fractions were assayed for presence of EGR proteins.