Skip to main content
Fig. 5 | Breast Cancer Research

Fig. 5

From: Novel mechanism for OSM-promoted extracellular matrix remodeling in breast cancer: LOXL2 upregulation and subsequent ECM alignment

Fig. 5

OSM-induced LOXL2 promotes ECM crosslinking and alignment of collagen I fibers. a Collagen contraction assay was performed using 1.5 mg/mL collagen I matrices or “pucks” seeded with MCF7 cells and treated for 48 h with OSM or OSM with βAPN (500 μM) or PXS-5120A (200 nM) [LOXL2 inhibitors]. After 48 h, dissecting microscope images of the collagen I matrix depict significantly more contraction in the OSM-treated samples, which is reversed in the presence of LOXL2 inhibitors. Scale bar = 2 mm. Graph quantifying the change in area of the matrix (in mm2) due to contraction. Collagen I fiber contraction correlates to fiber crosslinking and is reversed with the inhibition of LOXL2. b The same experiment as above was performed with MDA-MB-468 cells. Representative images after 48 h and accompanying graph are depicted using the same scale as above. As with MCF7 cells, LOXL2 inhibition prevented contraction due to OSM-induced LOXL2, but overall contraction is not as pronounced. Scale bar = 2 mm. c Confocal images of the collagen “pucks” depict the increase in collagen I fiber (green) alignment between MCF7 cells (red) in collagen I matrices that are treated with OSM for 36 h. Alignment is not observable with the addition of LOXL2 inhibition using βAPN (500 μM). Magnification × 63; scale bar = 50 μm. d Representative images with areas of greatest collagen I fiber density emphasized using ImageJ image processing. This clearly highlights the increase in fiber density and alignment present with OSM treatment, which reversed by LOXL2 inhibition. e Graph depicting the average fiber dispersion coefficient of collagen I fibers perpendicular to and bridging MCF7 cells in collagen I matrices. Confirms qualitative data that OSM treatment significantly increases alignment which is reversed with LOXL2 inhibition. (All experiments n = 3+; **p < 0.01, ***p < 0.001; Students t test and one-way ANOVA)

Back to article page