Journal: Biochemistry and Biophysics Reports

Article Title: Up-regulation of Syndecan-4 contributes to TGF-β1-induced epithelial to mesenchymal transition in lung adenocarcinoma A549 cells

doi: 10.1016/j.bbrep.2015.11.021

Figure Lengend Snippet: 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 . 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.

Article Snippet: Images of fluorescent staining were obtained for each field and merged by fluorescent microscope (Nikon Eclipse Ni-E).

Techniques: Knockdown, Expressing, Transfection, Isolation, Quantitative RT-PCR, Microscopy, Staining

Journal: Biochemistry and Biophysics Reports

Article Title: Up-regulation of Syndecan-4 contributes to TGF-β1-induced epithelial to mesenchymal transition in lung adenocarcinoma A549 cells

doi: 10.1016/j.bbrep.2015.11.021

Figure Lengend Snippet: SDC4 enhances TGF-β1 stimulated cellular restitution, chemotaxis and proliferation. A, Scratched restitution assay. Cells were seeded in 24-well culture plates at 2×10 4 cells/well. Forty-eight hours after transfection with indicated siRNA, a scratch was made using 200-μl micropipette tip. Upper: 0 and 20 h after wounding. Black dotted lines indicate the wound edge. Lower: Percentage (%) change in migration as determined by comparing the difference in wound width ( n =3). Each bar represents means ±SEM; ※, p <0.05. Scale bars: 100 μm. B, Transwell chemotaxis assay: The transfected A549 cells (2×10 4 cells) were loaded into 24-well inserts (8.0 μm-pore size) with DMEM medium containing 0.5% FBS in the presence or absence of TGF-β1 (5 ng/ml). Lower wells of the plate were filled with DMEM with 10% FBS. After 24 h, remove A549 cell on upper-side, then lower side of membrane were stained with DAPI. DAPI-positive migrated cells were counted with a fluorescent microscope. Each bar represents means ±SEM; ※, p <0.05. Scale bars: 50 μm. C, Cellular proliferation assay: The cellular proliferation was assessed with MTT assay. A549 cells were transfected with indicated siRNAs and reseeded into 96-well plate (5×10 3 cells/well) in the presence or absence of TGF-β1 (5 ng/ml), then after 72 h incubation, MTT assay was performed. Data represent means ±SEM; ※, p <0.05.

Article Snippet: Images of fluorescent staining were obtained for each field and merged by fluorescent microscope (Nikon Eclipse Ni-E).

Techniques: Chemotaxis Assay, Transfection, Migration, Pore Size, Membrane, Staining, Microscopy, Proliferation Assay, MTT Assay, Incubation