Figure 4

SPP1 knockdown impairs wound healing and cell invasion. Stable ST8814, S462, S1844.1 and S1507.2 cell lines, expressing either non-target or SPP1 shRNA, as indicated, were subjected to a wound healing assay. a) Pictures of cells were taken at 0 and 18 h after wounding. b) The percentage of cell migration was calculated using three individual experiments. SPP1 knockdown significantly reduced cell migration during wound healing in all four MPNST cell lines [P values for the four cell lines were as follows: ST8814 (0.03), S462 (0.001), S1844.1 (0.001), S1507.2 (0.003)]. c) SPP1 knockdown significantly reduced cell invasiveness; crystal violet was used to stain invasive cells. Cells were then eluted with 1% (w/v) SDS, and the absorbance was read at 550 nm on a Genova MK3 Lifescience Analyser; the number of cells was calculated by plotting the absorbance against a standard curve. The percentage of invasive cells was calculated in three individual experiments. SPP1 knockdown significantly reduced cell invasion in all four MPNST cell lines, suggesting a potential role for SPP1 in metastasis [P values for the four cell lines were as follows: ST8814 (0.0001), S462 (0.001), S1844.1 (0.0001), S1507.2 (0.001). *P < 0.05 when comparing treated vs. untreated cells].