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. 2017 Aug 2;8(39):66033–66050. doi: 10.18632/oncotarget.19789

Figure 5. Influence of ERα knockdown, methyl-β-cyclodextrin (MBCD), N-acetyl cysteine (NAC), PD98059, LY294002, an Akt inhibitor (AKI), and rapamycin on 7-ketocholesterol (7-KC)-reduced doxorubicin accumulation in MCF-7 cells.

Figure 5

Panel (A) shows the 7-KC-induced changes of doxorubicin accumulation in ERα siRNA- and scramble-siRNA-transfected cells. The lipid raft integrity was disrupted using 10 mM MBCD. After siRNA-transfection or MBCD pre-exposure, cells were exposed to 7-KC in the medium supplemented with FBS for 48 h and then cellular doxorubicin accumulation were determined. The results are presented as means ± SD of three independent experiments. *p < 0.05, compared to the vehicle control. Panels (C) and (D) show the effects of NAC, PD98059, LY294002, AKI, and rapamycin on the doxorubicin accumulation in MCF-7 cells cultured in media supplemented with 2% FBS and charcoal-stripped FBS (DCCFBS), respectively. Cells were pre-exposed to a scavenger or inhibitor at a concentration as indicated in the figure for 1 h prior to 48-h co-treatment with scavenger/inhibitor and 7.5 μM 7-KC. After co-treatment, cellular accumulation (1 h) of doxorubicin was determined using 3 μM doxorubicin as described in the Methods. The results are presented as means ± SD of three to four independent experiments. *p < 0.05, compared to vehicle control. #p < 0.05, the comparison between two treatments as indicated.