Figure 1

Manipulation of LOXL2 expression levels in human MCF10A normal mammary epithelial cells. (A) Western blot of secreted LOXL2 in CM generated from MDA MB-231 cells (MDA 231), MCF10A cells infected with LOXL2 (10A L2) and vector alone (10A cont) revealed that LOXL2 protein expression was upregulated in the 10A L2 cells to a level similar to that detected in MDA cells. β-Actin was used as a loading control. (B) Quantitative real-time PCR (qRT-PCR) of LOXL2 mRNA levels in manipulated MCF10A cells showed that LOXL2 mRNA levels were upregulated in 10A L2 cells. P = 0.009. (C) Morphologies of the manipulated MCF10A cells compared with WT cells showed that upregulation of LOXL2 did not produce significant alterations on cells plated on 2D tissue-culture plastic. Cells were viewed under a microscope (Leica DM1L), and representative images were taken. (D) 2D MTS proliferation assay of manipulated MCF10A cells suggested that when cultured on plastic, LOXL2 expression did not alter proliferation of the 10A cells. Error bars represent SEM for three independent experiments. (E) 3D MTS proliferation assay of manipulated MCF10A cells with manipulated LOXL2 expression plated within Matrigel suspension suggested that increased LOXL2 expression increases proliferation of the 10A cells in 3D. Error bars represent SEM for three independent experiments. P = 0.034 for day 6.