Figure 2
From: Characterization of extracellular matrix macromolecules in primary cultures of equine keratinocytes
Ln-332 synthesis and processing in equine keratinocytes. A. Immunofluorescence analysis of expression of Ln-332 α3β3γ2 and individual Ln-332 subunits in skin and lamellar keratinocytes. Scale bar = 10 μm. Inset in skin keratinocytes panel A demonstrates Ln-332 α3β3γ2 localization in sub confluent cells compared to confluent cells in larger image. Images of equine fibroblasts stained with each antibody serves as a negative control. As these cells do not produce Ln-332, nuclear staining with DAPI is shown in each panel. B. Immunoblots of individual Ln-332 subunit forms present in cell and matrix layers as well as secreted media from equine keratinocytes. Aliquots of acetone precipitated conditioned media, cell layers and matrix preparations were analyzed with each Ln-332 subunit antibody. Serum free conditioned media of the human squamous cell line SCC25 and the mouse epidermal cell lime Pam212 serve as positive controls for Ln-332 subunit molecular weight and processing. C. Confocal microscopy of equine keratinocytes grown on coverslips labelled for Ln-332 α3β3γ2 localization. Images are shown from both the basal and middle section of the cell monolayer. Scale bar = 10 μm.