Figure 5
From: The histone demethylase KDM4B regulates peritoneal seeding of ovarian cancer
KDM4B supports ovarian cancer cell invasion, migration and 3D spheroid formation. (a) SKOV3ip.1 Matrigel Invasion Assay. 5 × 104 SKOV3ip.1 cells expressing the indicated shRNA were seeded to Boyden chambers coated with Matrigel and cultured for 24 h in the indicated oxygen tensions. Cells were trypsinized and quantified as described in the Materials and methods. (b) Migration assay. 5 × 104 SKOV3ip.1 cells were seeded to empty Boyden chambers, cultured and quantified as in (a). Data in (a) and (b) represent mean±s.e.m. for five independent experiments (n=3). (c) Representative image of SKOV3ip.1 invasion assay. Filters treated in parallel to those in (a) were fixed and stained, as described in the Materials and methods. × 100 magnification, scale bar, 200 μm. (d) OVCAR8 invasion assay performed as in (a). (e) Migration assay for OVCAR 8 cells, treated as in (b). Data in (d) and (e) represent the mean±s.e.m. for four independent experiments, n=3. (f) Representative images of OVCAR8 Matrigel invasion assay. × 100 magnification, scale bar, 200 μm. (g) SKOV3ip.1 spheroid formation assay. SKOV3ip.1 cells transduced with the indicated shRNA were cultured in ULA dishes in 21% and 2% O2 as described in the Materials and methods. After 4 days, spheroid volume was calculated. Data represent mean volume±s.e.m. n=16. (h) Representative images (median volume) of SKOV3ip.1 spheroids. × 100 magnification, scale bar, 100 μm. (i) OVCAR8 spheroid formation assay, conducted as described for (g). (j) Representative images (median volume) of OVCAR8 spheroids. × 100 magnification, scale bar, 100 μm. For (a, b, d, e, g, i), significance was calculated relative to shGFP control at each oxygen condition, using Mann–Whitney U-tests (*P<0.05; #P<0.005).
