Wnt5a promotes the adhesion but inhibits the migration of hDPCs.

Abstract

<p>A: A total of 25,000 cells were seeded for the indicated times and nonadherent cells were rinsed off. Adherent cells were stained with crystal violet and were analyzed spectrophotometrically. The number of adherent cells is shown as mean±SD from three independent experiments, with the number of DMEM-treated cells set as 100%. B: Confluent monolayers of hDPCs were scratched with a pipette tip and cultured in different medium containing 5% FBS for 12 hr. The relative migration distance of the wound edge was shown as mean±SD of three independent experiments, with the migration distance of DMEM-treated cells set as 100%. Bars, 100 µm. C: HDPCs were plated on glass coverslips coated with type I collagen and cultured with different medium for the indicated times. For FACs immunostaining, anti-vinculin antibody was used, and for F-actin staining, rhodamine-phalloidin was used, arrowheads mark FACs. Bars,10 µm. D: Confluent hDPCs were incubated with Wnt5a for the indicated times and Western analyses were used to detect the expression of vinculin, phospho-paxillin, phospho-MLC, with GAPDH, total-paxillin and total-MLC as loading control. The relative protein expression at 0 min is designated 1.0. *<i>p</i> < 0.05, n=3.</p