Figure 2

Integrins mediate morphology and invasive qualities of monocultured and co-cultured cells. (A-A”) PC3 cells grown in the presence or absence of either α6, β1 or both inhibiting antibodies. (A”) F-actin staining of PC3 cells in 3D culture. (B-B”) HS5 cells grown in the presence or absence of either α6, β1 or both inhibiting antibodies. (B”) F-actin staining of HS5 cells grown in the presence of both α6 and β1 inhibiting antibodies with acini formation (filled arrowhead). (C-C”) Co-cultured cells grown in the presence or absence of either α6, β1 or both inhibiting antibodies. (C”) In the presence of both α6 and β1 inhibiting antibodies, HS5 cells (STRO-1; green fluorescence) localised to the outer edge (filled arrowhead), while the PC3 cells (Cell Mask blue positive; STRO-1 negative) resided in the centre of the spheroid mass (unfilled arrowhead). (D) Quantification of the number of proliferating cells in PC3, HS5 and co-cultured cells over a 9 day period. (E) Quantification of the percentage of HS5 and PC3 cells proliferating in co-culture over a 9 day period. (F-F’) EDU labelling (red fluorescence) of HS5 (STRO-1 positive; green fluorescence) and PC3 cells (CellMask blue positive; STRO-1 negative) in co-culture at day 9. (G) Quantification of the number of cells to invade in the presence and absence of integrin inhibitors for PC3, HS5 and co-cultured cells. (H). Quantification of the percentage of invaded HS5 and PC3 cells in co-culture. (**p < 0.01, ***p < 0.001). Error bars denote S.E.M. Scale bars = 40 μm.