Figure 1
APE inhibited cell growth and migration of MDA-MB-231 and MDA-MB-468 cells. (a) MDA-MB-231 (left) and MDA-MB-468 (right) cells were cultured for 24 and 48 h in medium supplemented or not with APE at the indicated concentrations. Cell viability was then assessed by MTT assay and expressed as a percentage of untreated cells. Values represent the mean ± SD of three independent experiments. (b) Representative phase-contrast microscopy images showing the wound closure process at three different time points in MDA-MB-231 (left) and MDA-MB-468 (right) cells incubated or not with APE at the indicated concentrations. Images in the panels are relative to a single field of view, taken as qualitatively representative of a given experimental condition. Representative time-lapse videos related to MDA-MB-231 (videos S1–S4) and MDA-MB-468 (videos S5–S8) cells are included in Supplementary Information. (c) Evolution in time of the wound area A, normalized to the value A0 at time 0, for MDA-MB-231 (left) and MDA-MB-468 cells (right) incubated or not (ctrl) with APE. The linear range of each data series was fit in order to measure the wound closure velocity α. Upper bar diagram shows the values of the wound area A normalized to the value A0 for treated cells at time tf, when the A/A0 value for control is 0.2. Lower bar diagram shows the values of α (h-1) calculated at time tf for the control and treated cells. The values reported in the histograms represent the mean from several independent fields of view. Standard error of the mean was calculated and one-tailed and heteroscedastic t-test were computed to verify the statistical significance of the differences with respect to control samples (*P < 0.05, **P < 1 × 10–3, ***P < 5 × 10–4, ****P < 5 × 10–5, *****P < 5 × 10–6, *******P < 5 × 10–8). (d) The levels of MMP-2 and MMP-9 in MDA-MB-231 (left) and MDA-MB-468 (right) cells treated with APE at the indicated concentrations for 48 h were measured by Western blot. All results were obtained from at least three independent experiments. α-tubulin and β-actin were used as standard for the equal loading of protein in the lanes. The full-length blots are included in Supplementary Information (Fig. S3).
