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. Author manuscript; available in PMC: 2020 Mar 1.
Published in final edited form as: Pharmacol Res. 2019 Jan 2;141:331–342. doi: 10.1016/j.phrs.2019.01.002

Fig. 3. BCRP protein expression and efflux activity in human BeWo trophoblast cells treated with anandamide.

Fig. 3.

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.