Figure 6. Skp2 promotes 3D culture and anchorage independence of growth in BKM-120 resistant cells.
(A) Representative images of MDA-MB-231 spheroids, from parental or BKM-120–resistant cells, that were infected with SKP2 shRNA lentiviral vector or control pLKO, and grown for 14 days in a 3-dimensional Matrigel/growth media mixture and treated with BKM-120 and/or MK-2206 (each 1 μM). Scale bar, 500 μm (4x magnification).
(B) Quantitation of spheroid sizes from (A) was performed using ImageJ software. Data are means ± S.E.M. ***P < 0.001 by a two-sided Student’s t-test.
(C) Representative images of MDA-MB-231 colony growth, from parental or BKM-120–resistant cells, that were infected with SKP2 shRNA lentiviral vector or control pLKO, and grown in an agar/growth media mixture for 28 days in the presence of DMSO, BKM-120 and/or MK-2206 (each 1 μM).
(D) Number of colonies in soft agar obtained in (C) was quantitated using MatLab software. Data are means ± S.E.M. **P < 0.01 by a two-sided Student’s t-test. All data and blots are representative of at least 3 independent experiments.