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. Author manuscript; available in PMC: 2009 Nov 15.
Published in final edited form as: Cancer Res. 2008 Nov 15;68(22):9338–9347. doi: 10.1158/0008-5472.CAN-08-0265

Figure 3. Luciferase reporter gene assays of OATP1A2 promoter fragments.

Figure 3

Luciferase reporter gene assay of subcloned OATP1A2 promoter fragments revealed increased activity of luciferase in T47-D cells transfected with the −5120 to −6600 OATP1A2 promoter fragment after treatment with rifampin (10μM) (3A). In silico analysis of the responsive fragment revealed several potential DNA motifs (insert). Subsequent mutation of those motifs revealed a loss of transactivation mediated by rifampin (3B) in HepG2 transfected with a construct containing a mutated DR4 motif. To validate the link between OATP1A2 transactivation by PXR a chromatin immunoprecipitation was performed using different anti-PXR antibodies (A-20 and N-16 supplied by Santa Cruz). The PCR detecting a DNA fragment which includes the identified DR4 motif suggests that this DNA motif is involved in the PXR mediated induction of OATP1A2 (3C). Data are expressed as mean ± SD; n=3, * p<0.05, t-test.