Association of DJ-1 and SREBP with the LDLR promoter.

<p>A. Chromatin immunoprecipitation assays were carried out using chromatin prepared from NIH3T3 (a) and D2 (b) cells. Chromatin was immunoprecipitated with anti-DJ-1, anti-SREBP-1 and anti-SREBP-2 antibodies or non-specific IgG. After extraction of DNA from precipitated chromatin, two regions spanning −180 to +54 and spanning −3920 to +54 were amplified by real-time PCR with specific primers and with amplified DNA as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038144#s4" target="_blank">Materials and methods</a>. Statistical analyses were carried out using Student’s <i>t</i>-test. Number of experiments (n) is 3. B. Gel-mobility shift assays were carried out using nuclear extracts from mouse liver and SH-SY5Y cells with IRDye800-labeled SRE oligonucleotide as a probe. a. NIH3T3 nuclear extracts were mixed with 50 and 100-times molar ratio of wild-type and mutated oligonucleotide compared to that of IRDye800-labeled SRE and subjected to gel-mobility shift assays. b and c. Mouse liver cell (b) or D2 cell (c) nuclear extracts were first reacted with the IRDye800-labeled SRE probe for 30 min at 0°C and then with an anti-DJ-1 antibody, anti-SREBP-1 antibody, anti-SREBP-2 antibody or IgG, and then separated on 4% polyacrylamide gel as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0038144#s4" target="_blank">Materials and methods</a>.