Fig. 4. Identification of nuclear factor binding sites by DNase I footprinting of the cyclin D3 promoter. A DNA fragment from the cyclin D3 gene promoter spanning sequences from −446 to −43 was end-labeled either at −43 (A and B) or at −446 (C). The fragment was incubated with 50 μg nuclear extract from Y10 cells cultured in the absence (lane 1) or presence (lane 2) of MGDF or with no nuclear extract (lane 3) and then digested with 1 U/mL (lane 1 and 2) or 0.1 U/mL (lane 3) of DNase. The protected sequences were identified by a sequencing reaction using primers end-labeled either at −37 or at −446. The nucleotide positions are indicated on the left. The sequences of the protected regions and nuclear factor putative binding sites are shown in (D). The data presented are of a representative experiment out of three performed.
Fig. 4.

Identification of nuclear factor binding sites by DNase I footprinting of the cyclin D3 promoter. A DNA fragment from the cyclin D3 gene promoter spanning sequences from −446 to −43 was end-labeled either at −43 (A and B) or at −446 (C). The fragment was incubated with 50 μg nuclear extract from Y10 cells cultured in the absence (lane 1) or presence (lane 2) of MGDF or with no nuclear extract (lane 3) and then digested with 1 U/mL (lane 1 and 2) or 0.1 U/mL (lane 3) of DNase. The protected sequences were identified by a sequencing reaction using primers end-labeled either at −37 or at −446. The nucleotide positions are indicated on the left. The sequences of the protected regions and nuclear factor putative binding sites are shown in (D). The data presented are of a representative experiment out of three performed.

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