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. 2009 Jun 24;284(33):22195–22205. doi: 10.1074/jbc.M109.028753

FIGURE 5.

FIGURE 5.

SREBP-1 and NF-YA bind to sequences in the promoter of the human LPIN1 gene. A, a radiolabeled DNA probe corresponding to nucleotides −2413 to −2404 of the LPIN1 promoter was incubated with nuclear extracts, with or without an excess of unlabeled probe corresponding to the LPIN1 SRE (LPIN1), the mutated SRE (LPIN1mut), or the identified SRE from the human LDLR gene (positive control). Supershift experiments were performed with anti-SREBP-1 or anti-glyceraldehyde-3-phosphate dehydrogenase (control) antibody. The closed arrowheads indicate the specific shifted bands. The experiments were repeated twice. B, a radiolabeled DNA probe corresponding to nucleotides −2386 to −2382 of the LPIN1 promoter was incubated with nuclear extracts, with or without an excess of unlabeled probe corresponding to the LPIN1 NF-Y-binding site (LPIN1), the mutated NF-Y-binding site (LPIN1mut), or the consensus binding site for NF-Y (Positive control). Supershift experiments were performed with anti-NF-YA or anti-PDI (Control) antibody. The closed and open arrowheads indicate the specific shifted bands and the supershifted bands, respectively. The experiments were repeated twice.