1. The subcellular localization, dimerization with Arnt and DNA‐binding activity of AhR are altered by the activation of Gα13.
The subcellular localization, dimerization with Arnt and DNA‐binding activity of AhR are altered by the activation of Gα13. (A) COS‐7 cells expressing HA–AhR, GST–Arnt or Gα13Q226L were stimulated with 3‐MC and/or LPA for 24 h. The localization of AhR was visualized by immunostaining with the HA antibody. (B) COS‐7 cells transfected as indicated were stimulated with 1 μM 3‐MC for 6 h. The protein complexes were precipitated and analysed by immunoblot analysis. (C) COS‐7 cells transfected with the indicated combinations of plasmids were stimulated with 1 μM 3‐MC for 6 h. Nuclear extracts were analysed by EMSA with the radioactively labelled AhR‐binding element DNA probe. An arrow indicates the AhR–Arnt complex binding to the probe. (D) Hepa1c1c7 cells were transfected with the indicated siRNA mixture, and stimulated with 1 μM 3‐MC for 12 h. The expression of CYP1A1 was analysed by the quantitative RT‐PCR method. Expression of AhR, AIP and β‐actin were analysed by immunoblot. 3‐MC, 3‐methyl cholanthrene; AhR, aryl hydrocarbon receptor; AIP, AhR‐interacting protein; Ctl, control; EMSA, electrophoretic mobility shift assay; GST, glutathione S‐transferase; HA, haemagglutinin; LPA, lysophosphatidic acid; RT–PCR, reverse transcription–PCR; siRNA, short interfering RNA.
Asuka Nakata et al. EMBO Rep. 2009;10:
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