Additional file 1.

Figure S1. Effect of Y477F ezrin on growth of AC2M2 cells in 3D Matrigel cultures. Panels A and B) AC2M2 cell clones expressing pCB6 empty vector or Y477F ezrin (clones A43 and C13) were cultured in 3D Matrigel. Representative phase contrast images of 9 day cultures of clones pCB6 (A) and C13 (B) photographed with a 4x objective are shown. Panel C) AC2M2 cell clones described above were cultured in 96 well plates (104 cells/100 μl/well) with 20% Matrigel, supplemented with Phenol Red-free complete DMEM medium. After 3 days, an MTT assay was performed according to the manufacturer's instructions. Values represent mean O.D. (Absorbance at 570 nm) of 4 wells +/- SD. No significant difference in growth was detected as determined by one way ANOVA (p = 0.319). Panel D) The number of colonies per well in 9 day cultures described in A and B was counted visually using phase contrast microscopy, and plotted as the mean of three wells +/- SD. The colony forming ability (% colonies per 7.5 x 103 cells plated) for each group is indicated. A marginal increase in colony forming ability was apparent in A43 and C13 (perhaps due to some clustering of more diffuse pCB6 colonies), but this difference was not significant as determined by one way ANOVA.

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Mak et al. BMC Cancer 2012 12:82   doi:10.1186/1471-2407-12-82