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. 2011 Jul 1;1(3):231–239. doi: 10.4161/spmg.1.3.18019

Figure 4.

Figure 4

PFOS inhibits RA-mediated transactivation of retinoic acid response element (RARE), as illustrated using luciferase reporter assay. The day before transfection, MCF7 cells were plated into 24-well tissue culture dishes at a density reaching 70–80% confluence by the time of transfection. Transfection was performed using LipofectAMINE™ 2000 reagent (Invitrogen) and OPTI-MEM®I medium (GIBCO) with 250 ng of retinoic acid reporter (Addgene). Six hours after transfection, the transfection medium was replaced by a complete medium and different doses of retinoic acid (10–1,000 nM) or PFOS (0.1–100 µM) or co-treatment of retinoic acid (1,000 nM) and PFOS (0.1–100 µM) were added. After 24 h incubation, the cells were then lysed in the passive lysis buffer. Firefly luciferase activities were measured using the Luciferase reporter assay system (Promega) and the multilabel reader VICTOR™X4 (PerkinElmer).