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Fig. S3
Blocking GJ communication inhibits endothelial cell-induced breast cancer and melanoma colony formation in vitro. (A) 4T-1 tumor cells expressing GFP (green) were cultured for 5 days in three-dimensional Culturex matrix alone or with human EA.hy926 endothelial cells (red, RFP transfected) and treated with vehicle (DMS0) or CBX (20 μM). Upper panel shows 4T-1 cells that were cultured alone and vehicle treated (days 1 and 5) or treated with CBX (day 5). Lower panel shows 4T-1 cells that were cocultured with EAhy.926 and vehicle treated (days 1 and 5) or treated with CBX (day 5). Note, that co-culture with EA.hy926 cells promotes 4T-1 tumor cell colony formation, while CBX treatment inhibits it. (B) Average size and number of 4T-1 or 4T-1KNcx43 tumor cell colonies that were cultured alone or with EA.hy926 cells and treated with CBX or vehicle (control) for 5 days. (C) b16 melanoma cells expressing GFP (green) were cultured for 5 days in 3D Culturex matrix alone or with human EA.hy926 (red, RFP) endothelial cells and treated either with the vehicle (DMSO) or CBX (20 μM). Upper panel shows b16 cells that were cultured alone and vehicle treated (days 1 and 5) or treated with CBX (day 5). Lower panel shows b16 cells that were co-cultured with EA.hy926 and vehicle treated (days 1 and 5) or treated with CBX (day 5). Note, that co-culture with EA.hy926 cells promotes b16 tumor cell colony formation, while CBX treatment inhibits it. (D) Average size and number of b16 or b16KNcx26 cell colonies cultured alone or with EA.hy926 and treated with CBX or vehicle (control) as above. Bars = 200 μm. * = P values < 0.05.