Biomedical Engineering Reference
In-Depth Information
Fig. 4.1 b
1
integrin expression correlates with collagen IV-cell adhesion and differentiation in
keratinocyte subpopulations a For cell adhesion assay, keratinocytes, with or without
preincubation with anti-b
1
integrin or anti-vimentin antibody (used as control) for 1 h at
37 C, were seeded onto microplate coated with or without collagen IV. The adhesion was
blocked 2 h later. The adhered cells were fixed with formaldehyde and stained with 0.2 % crystal
violet. b Protein extracts from keratinocyte subpopulations, isolated by their adhesiveness
capacity to collagen IV, were analyzed by western blotting for b
1
integrin expression. b-actin
expression was used as internal control. c Cells from KSC, TA and PM were fixed in situ in 4 %
paraformaldehyde and cytospun onto slides. After removal endogenous peroxidase activity, slides
were incubated with anti-involucrin antibody for 1 h. They were then incubated with secondary
antibody for 1 h followed by incubation with ABC complex for 1 h. Finally, 3-amino-9-
ethylcarbazole was used for visualization. Relative intensity of staining was quantified by
ImageJ64 software
integration of cell migration and keratinocyte proliferation as well as extracellular
matrix deposition, angiogenesis, and remodeling by fibroblasts that produce, among
other substances, agents promoting keratinocyte growth [
27
,
28
].
