Fig. 2

Podoplanin silencing reduces motility, migratory capacities and invasiveness of BcPAP and TPC1 cells. a. Wound healing motility assay. Representative light microscope images showing healing of wounds in monolayers of TPC1 and BcPAP cells transfected with siPDPN or control siNeg Magnification: 200x. b. Chamber migration assay. siPDPN or siNeg-transfected TPC1 and BcPAP cells were seeded in 8-μm Boyden insert chambers, with the lower reservoir filled with medium supplemented with 10% FBS as a chemoattractant. After 24 h, cells that had passed through the 8-μm membrane pores were fixed, stained with Diff-Quik and photographed at a 40x magnification.. Data are presented as means ± standard errors of the mean (SEM) of at least three independet experiments. c. Matrigel invasion assay. siPDPN- or control siNeg-transfected TPC1 and BcPAP cells were seeded in 8-μm Matrigel Invasion Chambers, with the lower reservoir filled with the culture medium supplemented with 10% FBS as a chemoattractant. After 24 h, cells that had passed through the 8-μm pores in the membrane were fixed with, stained with Diff-Quik Kit, and photographed at 40x magnification. The relative invasiveness of transfected cells is presented in graph form. Data are presented as the mean ± SEM of at least three separate experiments