Cell adhesion activities of purified laminin-332s. A, cell
adhesion activities of vector-Lm332 (crosses), GnT-III-Lm332
(open squares), and GnT-V-Lm332 (closed circles). The
96-well plates were coated with the indicated concentrations of each Lm332.
After 1.2% BSA blocking, Lm332-null keratinocytes suspended in supplement-free
keratinocyte medium were plated to each well and incubated for 20 min. Each
point represents the mean of triplicate assays. Other experimental conditions
are described in “Experimental Procedures.” B, 2 μg/ml
Lm332s were coated to the 96-well ELISA plates at 4 °C overnight. After
blocking those wells with 1.2% BSA, coating efficiency of three kinds of
Lm332s was estimated by ELISA using anti-laminin γ2 chain antibody.
C, inhibitory effects of function blocking antibody against integrin
α3(anti-α3) or control mouse IgG (IgG)
on cell adhesion of Lm332-null keratinocytes to different kinds of Lm332s. The
numbers in parentheses indicate the concentrations for Lm332
(μg/ml). Each bar represents the mean ± S.D. of triplicate
assays.