Fig. 4

PD0325901 significantly reduces aggregate spreading and ex vivo dispersal. Aggregates of GBM-3 cells were plated onto tissue culture plastic in complete medium with DMSO (a, c) or PD0325901 (b, d) and incubated for 24 h, whereupon they were fixed, permeabilized and stained with rhodamine-phalloidin. Low Mag (a, b) and High Mag (c, d) images were collected. Note single cell dispersal from untreated aggregates (a, c), in contrast to a higher level of cell-cell contact and actin stress fibers in response to PD0325901 treatment. (b, d). Scale bars in (a) and (c) are 100 μm. For GBM-1-3, PD0325901 decreased z-axis dispersal of GBM cells through a normal human astrocyte seeded 3D scaffold (e). Asterisks represent pair-wise comparison, Student t-test p < 0.05. No significant difference in z-axis dispersal was observed for GBM-4 (p = 0.9731)