A. Protein expression of BCRP in AEA-treated cells and corresponding densitometry analysis. Lysates were prepared from
control and AEA (10 μM, 24 h) treated BeWo cells and analyzed for expression of BCRP protein (72 kDa) by Western blotting.
β-Actin (42 kDa) was used as a loading control. B and C. Inhibition of BCRP Hoechst 33342 and zearalenone efflux activity
by AEA. Control and AEA (10 μM, 24 h) treated BeWo cells were trypsinized and subjected to either the Hoechst 33342 (5
μM, B) or zearalenone (50 μM, C) cell accumulation assay as described in the Materials and Methods. Data are
presented as mean ± SE (n = 3-5). D. Inhibition of BODIPY-glyburide transport across BeWo monolayers. Control and AEA (10
μM, 24 h) treated BeWo cells in transwell inserts were assessed for basolateral-to-apical translocation of BODIPY-glyburide
(1 μM) over 2 h as described in the Materials and Methods. Data represent the mean ± SE (n = 3 independent
experiments). One group of cells in each experiment were incubated in the presence Ko143 (1 μM) during the assay to provide
a positive control for BCRP inhibition. Asterisks (*) represent statistically significant differences (p < 0.05) compared
to control cells.