Fig. 4. (A) Expression and deposition of EMILIN1 and FN in vitro. Cells were grown in vitro for 48-72 hours, fixed and stained with anti-EMILIN1 or anti-FN antibodies. Bar, 25 µm. (B). Confocal images obtained from cryostat sections of first-trimester placenta using triple-labeling staining. Nuclei are visualized with Sytox Green and pseudocoloured in blue, EMILIN1 is detected with Alexa Fluor® 633 and pseudocolored in green, trophoblast cells are stained with anti-cytokeratin 7 (red, upper panel), and stromal cells with anti-vimentin (red, lower panel). EMILIN1 is deposited in proximity to stromal cells, whereas it seems absent near to trophoblast cells (magnification). Images were acquired with a Leica TCS SP2 confocal system (Leica Microsystems Heidelberg).