Journal: Laboratory investigation; a journal of technical methods and pathology
Article Title: Wnt Signaling Induces Epithelial-Mesenchymal Transition with Proliferation in ARPE-19 Cells upon Loss of Contact Inhibition
Figure Lengend Snippet: Contact inhibition is unlocked by EGTA only in the presence of EGF and/or FGF-2, but not TGF-β1. (a) Immunostaining of BrdU was performed in ARPE-19 cells cultured to day 7 post-confluence after being treated without or with 1mg/mL EGTA immediately followed by PBS, 10 ng/mL EGF, 20 ng/mL FGF-2, 10 ng/mL TGF-β1, 10 ng/mL EGF + 20 ng/mL FGF-2, or 10 ng/mL EGF + 20 ng/mL FGF-2 + 10 ng/mL TGF-β1 for 1 day. Without EGTA, no BrdU labeling was found. When EGTA was added, BrdU labeling was promoted by EGF, FGF-2, or EGF+FGF-2, but not by PBS, TGF-β1 or EGF+FGF-2+TGF-β1. Scale bar, 100 μm. (b) The BrdU labeling index was significantly increased by EGF, FGF-2, or in combination (highest) when EGTA was added (*, †, and ‡, P
Article Snippet: Antibodies and Reagents Dulbecco’s modified Eagle’s medium (DMEM), Ham’s/F12 medium, human epidermal growth factor (EGF), HEPES buffer, phosphate-buffered saline (PBS), amphotericin B, gentamicin, fetal bovine serum (FBS), and Alexa fluor-conjugated secondary IgG were purchased from Invitrogen (Carlsbad, CA).
Techniques: Inhibition, Immunostaining, Cell Culture, Labeling