Fig. 7.

Fyn is required for cellular migration, invasion and polarization in HEK293T cells and MSC. a Micrographs showing scratch healing after 16 h of incubation of the indicated cells with control, Wnt3a or Wnt5a-medium. Assays were performed as indicated in Methods. b Transfected MSC were seeded on Transwells and stimulated for 16 h with Wnt3a and Wnt5a when indicated. Cells that have migrated through the filter cells were fixed and stained with crystal violet, and optical density was quantified at 590 nm. Results are presented as mean ± range from two independent experiments. c, d, Transfected MSCs or HEK293T cells treated with control, Wnt3a or Wnt5a-conditioned medium were seeded in Transwell chambers containing 1 mg/mL collagen type I. After 16 h (c) or 36 h (d) of incubation, cells that have migrated through collagen were quantified a described. Results are presented as mean ± range from three independent experiments. e Polarized cortical actin was determined as indicated in Methods in MSC transfected with control, Fyn or Src shRNAs treated with Wnt5a when indicated. Cells were fixed and stained for F-actin and nucleus with Dapi. f The percentage of cells presenting polarized actin in e is shown. At least 100 GFP-positive cells were counted for each condition. Results are presented as mean ± SD from three independent experiments. *p < 0.05; **p < 0.01