Figure 1

LSD1 is induced in mesenchymal cells and promotes breast cancer EMT markers. (a) LSD1 transcript levels measured by qPCR in MCF-7 cells after incubation with PMA + TGF-β or withdrawal of PMA+TGF-β after 60 h incubation at the indicated time points. Data are expressed as fold change relative to 0 h stimulation or 0 h stimulation withdrawal (n = 3). *Indicates significance relative to 0 h stimulation. (b) Immunofluorescence microscopy was performed on cells fixed and probed with primary anti-LSD1 or anti-Snail antibodies and DAPI. Representative images for each dataset are shown. Graph represents the TNFI values for LSD1 measured using ImageJ to select the nucleus minus background (n > 50 individual cells). (c) The PCC was determined for LSD1 and Snail (n = 40 individual cells). −1 = inverse of colocalization; 0 = no colocalization; +1 = perfect colocalization. TCFI of E-cadherin and vimentin and TNFI of Snail and in MCF-7, MCF-7/PMA+ TGF-β and MDA-MB-231 cells after treatment with either (d) mock and LSD1 siRNA; or (e) vehicle alone and pargyline (n ≥ 30 individual cells). (f) LSD1 H3K4 demethylation activity assay was performed on MCF-7, MCF-7/PMA+ TGF-β, and MDA-MB-231 nuclear extracts in triplicate wells. LSD1 demethylase activity was calculated and graph depicts percentage of activity relative to MCF-7 cells (n = 2). All data represents the mean ±SE. Scale bars = 10 µm. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001, Mann-Whitney test.