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

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© The Author 2005. Published by Oxford University Press. All rights reserved

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Efficient binding of Pol II Δ5 to the c-myc promoter. (A) Raji cell lines carrying the episomal c-myc gene locus, and in addition (B) either LS*Mock (vector control), Pol II wt, or Pol II Δ5 were established. (C) The copy number of constructs was determined by Southern analysis with specific probes for the neomycin and hygromycin resistance genes. 1×, 10× and 100× correspond to 1, 10 and 100 copies/genome, respectively. (D) Proper expression of Pol II wt and Pol II Δ5 after removal of tetracycline was controlled by western blot analysis using anti-HA antibodies. Pol II Δ31 served as size control. (E) ChIP analysis revealed similar binding of Pol II Δ5 and Pol II wt to the c-myc promoter (lanes 3 and 11, respectively), while binding was not detectable in the Mock cell line (lane 7), or (F) to a non-transcribed region in the insulin gene.