Figure 4.

The inhibitors of importin (IMP) α/β mediated import importazole (IPZ) and ivermectin (IVM) reduce the nuclear accumulation of AHR. MCF-7^ΔAHR cells were transfected with EYFP-AHR^WT and treated with IPZ or IVM for 90 min at concentrations of 10 µM and 17.5 µM, respectively. Relative nuclear intensity of around 50 cells (sorted according to 0 min value) at the time of treatment (0 min) and after 90 min of treatment with IPZ (a) and IVM (b), respectively. Representative images of cells before and after incubation with 17.5 µM IVM for 90 min (c). Slopes of nuclear transition (d) and mean of time-lapse measurements (e) after incubation with 10 µM IPZ or 7.5 µM IVM for 22 h followed by co-treatment with 10 µM indirubin (IND) for 15 min. Data is expressed as mean + /− S.D. for 12 cells. Control implies to cells treated with ligand only. One-way ANOVA, Dunnett's post-test, ***p < 0.001, ****p < 0.0001. MCF-7^WT cells were treated with IPZ or IVM for 24 h at concentrations of 10 µM and 7.5 µM, respectively. Thereafter, cells were co-treated with 2.5 µM IND for 2 h. Relative CYP1A1 and CYP1B1 mRNA levels determined by qPCR (f). Values were standardized against HPRT and normalized to a sample treated with DMSO and ligand (mean + /− S.D.; one-way ANOVA, Dunnett’s post-test, **p < 0.01, ****p < 0.0001).