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. 2005 Sep 12;33(16):5139–5144. doi: 10.1093/nar/gki802

Figure 2.

Figure 2

Transcription of the c-myc promoter by recombinant Pol II. (A) Tetracycline was washed out from cell lines at time point 0 h to allow expression of Pol II wt and Pol II Δ5. α-amanitin was added 24 h later to achieve quantitative inhibition of the endogenous Pol II. After 16 h, transcription of the episomal c-myc was induced by treatment of cells with SoB for 14 h. (B) northern analysis of c-myc RNA (left hand site), discrimination of endogenous and episomal c-myc RNA by S1 analysis (right hand site). P1t/P2t mark RNA from the endogenous, translocated c-myc gene (lane 14), P1/P2 RNA from the episomal c-myc (lane 13). (C) Nuclear run-on activity of Pol II Δ5 and Pol II wt at the episomal c-myc promoter. Cells were treated as described in (A), but nuclei were isolated 24 h after addition of α-amanitin, cells were not induced with SoB. (A–J) Long antisense oligonucleotides (50 nt) covering the c-myc promoter region; 7SK: antisense oligonucleotide for Pol III transcribed 7SK gene; T7: T7 RNA polymerase transcribed, uniformely labelled RNA of the c-myc promoter region.