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. 2015 Sep 28;210(7):1165–1184. doi: 10.1083/jcb.201506041

Figure 1.

Figure 1.

GIV is required for the invasive growth pattern of MDA-MB-231 cells in 3D cultures. (A–C) GIV depletion impairs the proinvasive growth phenotype of MDA-MB-231 cells in Matrigel cultures. (A) MDA-MB-231 cells stably depleted of GIV by two independent shRNA sequences (GIV shRNA1 and GIV shRNA2) or expressing a control shRNA sequence were seeded on Matrigel and pictured by DIC microscopy after 7 d. Bottom panels correspond to a magnified view of areas inside the white boxes drawn on the upper panels. White arrowheads highlight stellate-shaped cells protruding from the acini toward the matrix. (B) The efficiency of GIV depletion (∼65% for shRNA1 and ∼95% for shRNA2 cells) was determined by immunoblotting (IB) using the indicated antibodies. (C) Quantification of the acini size (n = 3; 200 acini per experiment). Each dot is the size of one acini, and the horizontal line is the mean ± SEM (***, P < 0.001). (D and E) GIV depletion does not alter MDA-MB-231 cell morphology or growth on plastic. MDA-MB-231 cells stably depleted of GIV by expression of GIV shRNA2 or expressing a control shRNA were seeded on plastic dishes and grown in complete media for 4 d. A representative field of these cells was pictured by DIC microscopy (D), and cells were counted every day using a hemocytometer (E). Results are depicted as mean ± SEM (error bars; n = 3).