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. 2010 Jul;177(1):119–131. doi: 10.2353/ajpath.2010.090772

Figure 4.

Figure 4

EMSAs detailing AP-1 activation in pBSMC on treatment with PDGF. Nuclear extracts prepared from control or PDGF-treated pBSMC were used in an EMSA and assessed for binding to a radiolabeled oligonucleotide carrying a consensus AP-1 binding sequence motif. Increased AP-1-DNA complex formation was detected in PDGF-treated pBSMC (A, compare lanes 2 and 5), and the complex was supershifted (SS) in the presence of antibody to (A) phospho-c-jun (lanes 4 and 7) or (B) Fra1 (lanes 3 and 5). An isotype-matched IgG control (A, lanes 3 and 6) did not affect mobility of the AP-1-containing complex. C: The PDGF-stimulated AP-1-DNA complex in pBSMC nuclear extracts (lane 2) was specifically competed out by increasing concentrations (25-, 50 -,and 100-fold molar excess) of unlabeled wt AP-1 consensus oligonucleotide (lanes 3–5) but not by an oligonucleotide carrying a mutation in the AP-1 binding motif (lanes 6–8). NS, nonspecific; FP, free (unbound) probe.