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. 2001 Apr 15;29(8):1672–1682. doi: 10.1093/nar/29.8.1672

Figure 8.

Figure 8

Tetracycline inducible expression of TBP-DR2 in HeLa cells. (A) Western blot analysis. Cellular extracts from uninduced (lane 1) and doxycycline-induced cells (lane 2) of one of the selected clones were probed with anti-histidine antibodies. Recombinant TBP-DR2 with a histidine tag (10 ng; lane 3), 25 µg HeLa whole-cell extract (WCE; lane 4) and 10 ng recombinant TBPwt without histidine tag (lane 5) served as controls. (B) Functional investigation of TBP-DR2 expressed in HeLa cells. To analyse whether the expressed TBP-DR2 was functionally active, 50 µg S100 from HeLa cells expressing TBP-DR2 was used to transcribe the PDR2 promoter (lanes 4–6) in vitro before (lane 5) and after (lane 6) doxycycline induction of these HeLa cells. Additionally, 50 µg S100 from untransfected HeLa Tet-on cells was used as a negative control (lane 4). As a control, all three extracts were simultaneously analysed by in vitro transcription of pUhU60.35 (lanes 1–3) and pUVAI DNA (lanes 7–9).