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. 2003 Jul 1;17(13):1617–1629. doi: 10.1101/gad.1097603

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Copyright © 2003, Cold Spring Harbor Laboratory Press

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Figure 2A. Characterization of genomic MMTV promoters. (A) Schematic illustration of MMTV promoter configuration in the 3134 cell genome. Cell line 3134 contains ∼200 copies of an MMTV-LTR-ras-BPV promoter construct stably integrated into Chromosome 4 in a perfect head-to-tail array. The 2400-bp MMTV promoter fragment characterized in these experiments was generated by KpnI digestion of isolated 3134 nuclei as described in Materials and Methods.

Figure 2B. Characterization of genomic MMTV promoters. (B) Multigel electrophoresis of a KpnI-digested low-salt nuclear extract prepared from untreated cells. The individual running gels ranged from 0.2% to 1% agarose and were poured in increments of 0.1%. After electrophoresis, specific MMTV promoter bands were detected by Southern blotting as described in Materials and Methods. Arrowheads indicate the center position of the promoter bands referred to as I and II in the text.

Figure 2C. Characterization of genomic MMTV promoters. (C) DEX-induced SacI cleavage of genomic MMTV chromatin. The 3134 cell nuclei isolated before (left lane, Uncut) and after exposure of cells to DEX for 1 h (right lane, Cut) were exposed to SacI as described in Materials and Methods. The 252-bp fragment represents the uncut promoter region. The 136-bp fragment is derived from the 252-bp fragment as a result of SacI restriction.

Figure 2D. Characterization of genomic MMTV promoters. (D) Multigel electrophoresis of a KpnI-digested low-salt nuclear extract prepared from DEX-treated cells. The 3134 cells were exposed to DEX for 1 h prior to preparation of the nuclear extract. The individual running gels ranged from 0.2% to 1% agarose and were poured in increments of 0.1%. After electrophoresis, specific MMTV promoter bands were detected by Southern blotting as described in Materials and Methods. Closed arrowheads indicate the center position of the promoter bands referred to as I_D, II_D, and III_D in the text. The asterisk and open arrowheads indicate the position of a fourth band that was not reproducibly present in the extracts.

Figure 2A. Characterization of genomic MMTV promoters. (A) Schematic illustration of MMTV promoter configuration in the 3134 cell genome. Cell line 3134 contains ∼200 copies of an MMTV-LTR-ras-BPV promoter construct stably integrated into Chromosome 4 in a perfect head-to-tail array. The 2400-bp MMTV promoter fragment characterized in these experiments was generated by KpnI digestion of isolated 3134 nuclei as described in Materials and Methods.

Figure 2B. Characterization of genomic MMTV promoters. (B) Multigel electrophoresis of a KpnI-digested low-salt nuclear extract prepared from untreated cells. The individual running gels ranged from 0.2% to 1% agarose and were poured in increments of 0.1%. After electrophoresis, specific MMTV promoter bands were detected by Southern blotting as described in Materials and Methods. Arrowheads indicate the center position of the promoter bands referred to as I and II in the text.

Figure 2C. Characterization of genomic MMTV promoters. (C) DEX-induced SacI cleavage of genomic MMTV chromatin. The 3134 cell nuclei isolated before (left lane, Uncut) and after exposure of cells to DEX for 1 h (right lane, Cut) were exposed to SacI as described in Materials and Methods. The 252-bp fragment represents the uncut promoter region. The 136-bp fragment is derived from the 252-bp fragment as a result of SacI restriction.

Figure 2D. Characterization of genomic MMTV promoters. (D) Multigel electrophoresis of a KpnI-digested low-salt nuclear extract prepared from DEX-treated cells. The 3134 cells were exposed to DEX for 1 h prior to preparation of the nuclear extract. The individual running gels ranged from 0.2% to 1% agarose and were poured in increments of 0.1%. After electrophoresis, specific MMTV promoter bands were detected by Southern blotting as described in Materials and Methods. Closed arrowheads indicate the center position of the promoter bands referred to as I_D, II_D, and III_D in the text. The asterisk and open arrowheads indicate the position of a fourth band that was not reproducibly present in the extracts.