Figure 5

AF induces cell cycle alterations in MDA-MB-468shAhR and Cal51shAhR. (A) MDA-MB-468shAhR cells were pretreated with 750 ng/mL Dox or an equivalent amount of vehicle for seven days to induce AhR knockdown. Cells were then treated with 0.1% DMSO or 25nM AF, with or without co-treatment with 750 ng/mL of Dox, for the corresponding length of time. DNA content was evaluated using propidium iodide staining. A representative graph of DNA content versus cell number is shown for DMSO control (top left panel) and for the accumulation of cells in S phase (top right panel). All data in the top panels is presented as percentage of total cells in G0/G1, S, and G2/M phase for each treatment, shown with standard deviation. Statistical analysis in the form of Student’s T-test was used to compare the percentage of S phase cells between 0.1% DMSO-treated and AF-treated cells was performed, but not labeled due to the stacked nature of the graph (B) Cal51shAhR cells were pretreated with 750 ng/mL Dox or an equivalent amount of vehicle for seven days to induce AhR knockdown. Cells were then treated with 0.1% DMSO or 250nM AF, with or without co-treatment with 750 ng/mL of Dox, for the corresponding length of time. DNA content was evaluated using propidium iodide staining. A representative graph of DNA content versus cell number is shown for DMSO control (top left panel) and for the accumulation of cells in S phase (top right panel). All data in the right panel is presented as percentage of total cells in G0/G1, S, and G2/M phase for each treatment, shown with standard deviation. Statistical analysis in the form of Student’s T-test was used to compare the percentage of S phase cells between 0.1% DMSO-treated and AF-treated cells was performed, but not labeled due to the stacked nature of the graph.