Fig. 2

DNMTi and HDACi reverse bsMCF cells malignant behavior in collagen and inhibit colony formation. a Representative images of 3D culture in bovine type I collagen. Images of structures formed in collagen were acquired after 6 days of culture. Magnification, 40×. Scale bar, 500 μm. b-c Quantification of the number of masses formed in collagen. d Classification of the masses. Type 1 masses show smooth outer surface and no protrusions present; type 2 masses show rough outer surface with 1–5 protrusions; type 3 masses show large amount of protrusions (> 5), indicate more invasive phenotype. Magnification, 40× for duct, and 100× for type 1–3 masses. e-f Quantification of the types of the masses. DAC, SGI, and MS275 treatment increase the number of masses with fewer protrusions. g Representative images of colony formation in agar methylcellulose matrix. Images were acquired after 9 days of culture. Magnification, 2×. Scale bar, 2 mm. h-i Quantification of the number of the colonies formed in methylcellulose matrix. *indicates p < 0.05. ** indicates p < 0.01 compared to control