Phenotypic analysis of stromal cells from 14 dpc murine fetal liver lines treated or not with OSM.

(A) Western blots for CK-8 and ASMA of AFT024 cells cultured in the absence or in the presence of OSM for 5 days. (B) Immunofluorescence studies of AFT024 cells treated or not with OSM. Cells were permeabilized and double-stained with mouse anti–CK-8 or –CK-18 (green) and mouse anti-ASMA–Cy3 (red). At day 5 and 10 there is no ASMA labeling. Scale bar = 10 μm. (C) FACS analysis of 3 stromal cell lines treated or not with OSM. Cells were analyzed for expression of intracellular components ASMA, CK-8, albumin, and AFP and of the cell surface marker E-cadherin. The histograms compare the levels of each marker in the presence or absence of OSM. Each panel represents an overlay of the fluorescence signal obtained with untreated (dark histogram) or treated cells (hatched histogram). Open histogram represents background staining with appropriate isotype control antibodies. A representative example of 3 experiments is shown. The arrows pointed down indicate down-regulation; arrows pointed up, up-regulation. Fluorescence intensity is indicated on the x-axes (C-D). (D) AFT024 cells treated or not with OSM were analyzed as described in panel C for the expression of other mesenchymal markers: osteopontin (OSP), calponin, collagen I (COLL I), EDa fibronectin (FN3E2), thrombospondin (TSP), and Stro-1 antigens.