Fig. 2

IL-32θ reduces breast cancer EMT, migration, invasion, and pro-malignancy factors in the CM treatment. a Constitutive expression system of 6x Myc-tagged IL-32θ in MDA-MB-231 cells by western blot and RT-PCR. b Schematic of in vitro model using MDA-MB-231-EV and MDA-MB-231-IL-32θ cells treated with CM from THP-1-derived macrophages. c Cellular morphological change in MDA-MB-231 EV and MDA-MB-231-IL-32θ cells in the absence (upper panel) or presence (lower panel) of CM. d mRNA expression levels of pro-malignancy factors in breast cancer cells were determined by real-time PCR (n = 5). e COX-2 and E-cadherin protein expression was analyzed by western blotting. f MMP-9 expression was detected by zymography. g Protein secretion levels of GM-CSF were measured by ELISA (n = 3). h and i The invasion or migration abilities of cells were performed using Matrigel-coated or non-coated transwell chambers. Related images were obtained from an upright microscope. j Migration or invasion intensities were quantitated based on an OD at 620 nm (n = 3). Scale bar, 10 μm (c); 100 μm (h); and 200 μm (i). All data are presented as the mean ± SEM and are analyzed using the Student’s t-test: *, p < 0.05. Western blot or RT-PCR or zymography images are the representative results of three independent experiments