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. Author manuscript; available in PMC: 2017 Oct 1.
Published in final edited form as: Placenta. 2017 Apr 12;55:29–36. doi: 10.1016/j.placenta.2017.04.006

Fig 6. Effects of modulating cholesterol on BCRP transporter activity in BeWo cells.

Fig 6

Cholesterol content in BeWo cells was modulated by treatment with MβCD (1 h, 5 mM) or pravastatin (PRAV, 48 h, 200 μM). Cholesterol repletion groups were treated with cholesterol-loaded MβCD (30 min, 5 mM) immediately before the experiment. A. Effects of MβCD and pravastatin on BeWo cell cholesterol content. Cells were treated with MβCD or pravastatin, lysates prepared and cholesterol content measured using an Amplex Red assay (n = 3 samples). B. Effects of modulating BeWo cell cholesterol on BCRP efflux transporter activity using Hoechst 33342 retention. Efflux transporter activity was measured by the ability of cells to retain the BCRP substrate Hoechst 33342. The BCRP substrate was quantified in relative fluorescence units (RFU) using a Nexcelom Cellometer and expressed as mean ± SD (n = 3 independent experiments). C. Effect of modulating BeWo cell cholesterol on BCRP efflux transporter activity using zearalenone retention. Efflux transporter activity was measured by the ability of the cells to retain the BCRP substrate zearalenone. Black bars represent cells where cholesterol was not restored. Grey bars represent cells where cholesterol was restored. Data are presented as mean ± SE (n = 4–6). Asterisks (*) represent statistically significant differences (p < 0.05) compared to control cells where cholesterol was not restored. Daggers (†) represent statistically significant differences (p < 0.05) compared to cells with the same treatment (CTR, MβCD or PRAV) where cholesterol was not restored.