Figure 2. Avicin D binds to GR and translocates it into the nucleus.

(A) A549 cells were incubated with 25 nM [³H] Dex in the presence or absence of 0–500 fold excess of cold Dex or cold avicin D. Following one hour incubation at 37°C, cells were lysed and radioactivity measured as described in the methods. (B & C) A549 cells were treated with avicin D (1 µM) for 0–60 min, or with Dex (1 µM) for 60 min at 37°C. (B) At the end of the incubation, cells were fixed and immunostained as described in the methods. (C) Western blot analysis of the nuclear extracts of treated cells was performed using anti- GR antibody as described in the methods. Actin expression was used as a loading control.