Effects of SDC4 knockdown on the expression of EMT-related genes and cell morphology. A, B, C, Cells were transfected with indicated siRNAs and after 48 h mRNA was isolated with the RNeasy Mini Kit (Qiagen), then real-time RT-PCR for Snail, Slug and E-cadherin was performed by indicated primers in Supplemental Table S1. Each bar represents means ±SEM (n=3). “※” indicates statistically significant (p<0.05). D, The cellular morphological images were observed by optical microscope. A549 cells were transfected by indicated siRNA(s), then exposed TGF-β1 for 48 h. Scale bar: 100 μm. E, Representative cellular images of A549 cells subjected to immunofluorescent staining. F-actin was stained with phalloidin. Images of fluorescent staining were obtained for each field and merged by fluorescent microscope (Nikon Eclipse Ni-E). A549 cells were transfected by indicated siRNA(s), then exposed TGF-β1 for 48 h. The blue color indicates nuclei, stained with DAPI. Scale bar: 50 μm.