Figure 7.

Members of the C/EBP family are not involved in the regulation of rhoB. (A) For gel retardation analysis the ³²P-labeled oligonucleotide derived from the sequence harboring the CCAAT box essentially required for activation of the rhoB promoter by genotoxic stress was used (NF-Y^rhoB, 5′-GGCTTCCCATTGGGTGGCTAT-3′). Competition experiments were performed using 20-fold molar excess of either non-labeled NF-Y^rhoB oligonucleotide (specific competition) or an oligonucleotide containing the consensus sequence for binding of C/EBP (C/EBP^a, 5′-TGCAGATTGCGCAATCTGCA-3′) (sc-2525; Santa Cruz). SComp, specific competition. (B) For band shift analysis the ³²P-labeled C/EBP-specific oligonucleotide (C/EBP^a) described in (A) was used. Competition experiments were done as described using either unlabeled C/EBP^a (SComp) or NF-Y^rhoB-specific oligonucleotide. SComp, specific competition. (C) A second consensus sequence described to bind C/EBP proteins (26) was used for band shift analysis (C/EBP^b, 5′-ATTCAATTGGGCAATCAG-3′). Specific competition (SComp) and competition with NF-Y^rhoB-specific oligonucleotide was done as described before.