Figure 8. Podocalyxin expression on lung endothelial cells is required for efficient spreading on laminin and collagen I matrices.

(A) Static adhesion of lung mECs isolated from Podxl^F/F (black bars) and Podxl ^ΔEC (white bars) mice to surfaces coated with fibronectin, laminin, gelatin, collagen I, and collagen IV. Primary mEC were plated on matrix-coated plates for 90 min, washed and adhesion quantified by crystal violet absorbance. The average absorbance for the uncoated wells was normalized to 1 and the relative absorbance (proportional to adhesion) is shown. (B) Spreading of lung mECs isolated from Podxl^F/F (black bars) and Podxl ^ΔEC (white bars) mice. Primary mECs were plated on matrix-coated transwells (0.4 µm pore size) and cultured for 48 h. The % monolayer coverage (spreading) was assessed by the threshold area of the cell monolayer via ImageJ on at least 3 independent cultures per genotype. The mean adhesion reported here was pooled from two independent experiments. (C) Representative bright field micrographs of spreading assay described in (E). Error bars  =  SEM, *Significantly different with P<0.05 by Student's t test or one-sample.