Technical Reference #1768

1768.

Galectin-3 promotes lamellipodia formation in epithelial cells by interacting with complex N-glycans on a3B1 integrin Chandrassegar Saravanan, Fu-Tong Liu, Ilene K. Gipson, Noorjahan Panjwani, Tufts University School of Medicine, Journal of Cell Science, 122(1768), (2009)
Link To Paper

Keywords:
Galectin-3, 3β1 integrin, Lamellipodia, Cell migration, Epithelium

Materials & Methods:
Studies were carried out using telomerase reverse transcriptase-immortalized human corneal epithelial (HCLE) cells (Gipson et al., 2003). The HCLE cells were maintained in keratinocyte serum-free medium supplemented with 0.2 ng/ml epidermal growth factor (EGF), 25 g/ml bovine pituitary extract (K-SFM; Invitrogen, Carlsbad, CA) and 0.4 mM CaCl2 at 37°C in a 5% CO2 incubator. For comparative purposes, primary cultures of human corneal epithelium and a skin keratinocyte cell line, HaCaT, were also used. Primary human corneal epithelial (HCE) cells were cultured from donor corneal rims of transplant corneas using endothelium-free explants (Ebato et al., 1988). HaCaT cells (a kind gift from Jonathan Garlick, Tufts University, Boston, MA) are spontaneously immortalized, nontransformed epithelial cells derived from adult human skin and are similar to normal skin keratinocytes (Boukamp et al., 1988). The HaCaT cells were grown in Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen) containing 5% fetal calf serum and 10 mM HEPES buffer.

Microscopic Technique
Fluorescent Microscopy

Cell Type(s)
HCLE cells