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. 2012 Nov 23;41(2):764–774. doi: 10.1093/nar/gks1120

Figure 2.

Figure 2.

p50S329D has reduced binding affinity for −1C-bearing κB-sites. (A) Competition binding analysis of p50wt (upper) and p50S329D (lower). Quantitative EMSA was performed with 32P-labeled oligonucleotide probe containing the −1A site (GGGAATTTCC) and a constant amount of recombinant p50. Increasing concentrations of non-labeled −1A, −1C (GGGACTTTCC) or non-specific DNA was added as indicated (see ‘Materials and Methods’ section). The amount of bound probe is plotted as a percent of control samples without competitor DNA. Data represent mean value ± SD of three separate experiments. (B) Scatchard plot analysis of p50wt (upper) and p50S329D (lower) binding affinity for −1C and −1A sequences. Quantitative binding analysis was performed using various probe concentrations and a constant amount of recombinant p50 (see ‘Materials and Methods’ section). Samples were analyzed by EMSA.