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Figure 2 | Molecular Cancer

Figure 2

From: Reduced annexin A6 expression promotes the degradation of activated epidermal growth factor receptor and sensitizes invasive breast cancer cells to EGFR-targeted tyrosine kinase inhibitors

Figure 2

Depletion of AnxA6 in invasive breast cancer cells promotes cell proliferation but inhibits cell motility. (A) AnxA6-depleted BT-549 cell lines were generated as described in “Materials and methods”. Whole cell lysates from the selected clones designated A6sh2 and A6sh5, were analyzed by Western blotting. (B) Cell proliferation assays. Cells were plated in quadruplicate in 24 well plates and cultured for 72 h in complete DMEM/F12 medium. Cell growth and viability was assayed using PrestoBlue reagent. Bars represent cell growth ± S.E.M from three independent determinations. * p < 0.05; *** p < 0.001. (C) Migration assays. Serum-starved AnxA6-depleted BT-549 and control cells were plated in duplicate in the upper chambers of 8 μm culture inserts. Complete DMEM/F12 was used as the chemoattractant. Bars represent the number of migrated cells/field ± s.d from three independent determinations. * p < 0.001. (D) Anchorage-independent growth of AnxA6-depleted BT-549 cells. Control (BT-EV) and AnxA6-depleted (BT-A6sh5 and BT-A6sh2) cells were maintained in matrigel 3D cultures for up to 10 days. Digital images of the colonies were captured with a digital camera (x10 magnification). (E-H) Analyses of AnxA6-depleted MDA-MB-231 cells. The expression levels of AnxA6 (E), Densitometric analysis of AnxA6 expression (F), cell migration (G) and growth in 3D cultures (H) were examined as described in A-D above.

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