Fig. 2

PLOD2 knockdown attenuates breast cancer cell migration in vitro. a PLOD2 was knocked down using two independent short hairpin RNAs (SHC and SHD) in MDA-MB-231 (MB-231) cells. qRT-PCR and Western blotting were used to detect PLOD2 expression in scramble (SCR) and PLOD2-knockdown cells (SHC, SHD). Error bars represent means ± SD. ^**P < 0.01. b Scramble (SCR) and PLOD2-knockdown cells (SHC, SHD) were cocultured in the presence or absence of adipocytes. After 3 days, cells were used for migration assays. Error bars represent means ± SD. ^**P < 0.01. Scale bars, 100 μm. c-d Scramble (SCR) and PLOD2-knockdown cells (SHC, SHD) were evaluated by migration (c) and 3D matrix invasion assays (d). Error bars represent means ± SD. ^**P < 0.01. Scale bars, 100 μm. e Expression levels of E-cadherin, N-cadherin, Vimentin, ZO-1 and Slug were detected by Western blotting in scramble (SCR) and PLOD2-knockdown cells (SHC, SHD). Immunofluorescence staining for E-cadherin (red) and N-cadherin (red) in scramble (SCR) and PLOD2-knockdown cells (SHC, SHD). Nuclei were stained with Hoechst (blue). Scale bars, 20 μm. f Immunofluorescence staining for F-actin (red) in scramble (SCR) and PLOD2-knockdown cells (SHC, SHD). Nuclei were stained with Hoechst (blue). Scale bars, 20 μm