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. Author manuscript; available in PMC: 2010 Apr 28.
Published in final edited form as: Curr Biol. 2009 Apr 2;19(8):688–693. doi: 10.1016/j.cub.2009.02.055

Figure 2. Ubx and Abd-B genes show promoter-proximal stalling of RNA Pol II in wing and imaginal discs tissues.

Figure 2

The Pol II ChIP was performed on wing and haltere imaginal discs, followed by conventional PCR and QPCR analysis. IP was done using a cocktail of antibody recognizing CTD of Pol II (8WG16) and Ser5 phosphorylated CTD of Pol II (H14) as well as each of them alone. (A) In the haltere discs Ubx gene is active and Pol II shows signals along the promoter (row 1) as well as exon regions (row 2), while in the wing discs it shows signal at the promoter region (row 4) and no signal at the exon (row5). Thus, supporting the fact that even when Ubx is inactive in the wing discs it still shows Pol II binding at the promoter regions and may be stalled/paused. The hsp70 gene shows Pol II signals at both haltere discs (row3) and wing discs (row 6). The abd-A promoter does not show Pol II signal in wing disc and was used as negative control (row 7) and rp49 was used as loading control (row8). (B) QPCRs using cocktail of Pol II antibody (8WG16 and H14) reveal signals of Pol II on Ubx promoter in wing discs, while haltere discs show similar signals of Pol II in promoter and exon regions. (C) Abd-B gene is known to be inactive in haltere as well as wing discs. QPCRs display enrichment of Pol II binding at the proximal-promoter region when compared to the exon in wing as well as haltere imaginal discs.