Technical Reference #1989

1989.

TGF-B1 + EGF-Initiated Invasive Potential in Transformed Human Keratinocytes Is Coupled to a Plasmin/MMP-10/MMP-1–Dependent Collagen Remodeling Axis: Role for PAI-1 Cynthia E. Wilkins-Port; Qunhui Ye; Joseph E. Mazurkiewicz; and Paul J. Higgins, Albany Medical School, Cancer Research, 69(1989), (2009)
Link To Paper

Keywords:
epithelial-to-mesenchymal transition; collagenolysis; MMP-10; PAI-1; keratinocyte

Materials & Methods:
Microscopy.HaCaT II-4 cellswere seeded onto collagen-coated coverslips (50 Ag/mL) or onto collagen gels polymerized in MatTek glass-bottomed dishes. Following treatment cells were fixed in 3% paraformaldehyde permeabilized blocked and incubated with primary and secondary antibodies for 1 h each. Coverslips were mounted using ProLong Gold with 4¶6-diamidino-2-phenylindole and viewed on an Olympus BX61 microscope with Image-Pro Lab software version 3.6.5. or an Olympus IX70 inverted scope with Image-Pro Plus software. For visualization of collagen digestion cells were seeded onto coverslips coated with collagen type 1 (50 Ag/mL) + DQ FITC-labeled collagen type 1 (25 Ag/mL).

Microscopic Technique
Immunofluorescence microscopy

Cell Type(s)
HaCaT II-4 cells