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. 2019 Apr 24;116(20):9959–9968. doi: 10.1073/pnas.1820604116

Fig. 4.

Fig. 4.

Compound 10 inhibits formation of the quaternary NFAT:Fos:Jun:DNA complex on the ARRE-2 site, but not on the GM-330 element, without inhibiting the direct binding of NFAT or AP-1 to DNA. 500 nM of Fos and 500 nM of Jun proteins were incubated with 20,000 CPMs of the indicated [γ32P] ATP-labeled oligos in the absence or in the presence of Compound 10 (concentrations: 250, 125, 62.5, 31.3, 15.6, 7.8, and 3.9 μM, respectively) for 10 min, then 10 nM of wild-type NFAT DBD or NFAT-RIT DBD was added for a further 20 min. DNA–protein complexes were analyzed by EMSA and are indicated by the arrows. Results are representative of at least two independent experiments. (A) Compound 10 inhibits formation of the quaternary NFAT:Fos:Jun:DNA complex on the murine ARRE-2 element. (B) Compound 10 does not inhibit binding of AP-1 (Fos:Jun heterodimers) to the consensus AP-1 binding site. (C) Compound 10 does not inhibit binding of NFAT monomers or dimers to the TNF κ3 element. (D) Compound 10 does not inhibit formation of the quaternary NFAT:Fos:Jun:DNA complex on the GM-330 GM-CSF enhancer element.