Fig. 7

Cholesterol depletion but not MMP inhibition reduces the 3D invasion of the four breast cancer spheroids. T-47D, HCC1954, Hs-578T and BT-549 spheroids were grown and treated exactly as in Fig. 6 and then tested for protrusion length and 3D invasion into 2 mg/mL Matrigel. A Representative phase contrast images at day 3 of invading Hs-578T and BT-549 spheroids, treated or not with mβCD (chol depletion) or GM6001 (MMP inhibition). Arrowheads, spheroid protrusions. B, C Time-course quantification of the mean protrusion length of Hs-578T (B) and BT-549 (C) spheroids embedded in Matrigel and treated or not (circles) with mβCD (squares) or GM6001 (triangles). Graphs are representative of 1 experiment with 5 spheroids per condition (in total 8–18 spheroids from 2–4 independent experiments). D–G Quantification of mean protrusion length at day 3 (D, F) and invasion in 3D at the last day of analysis (E, G) of cell lines treated with mβCD or GM6001. 3D invasion potential in E, G was obtained by subtracting the spheroid area increase without Matrigel (Fig. 6I-L) from the area increase with Matrigel (Fig. 6E-H). Each data point represents 1 spheroid (in total 3–15 spheroids from 2–5 independent experiments). Wilcoxon signed-rank test