Figure 5

Freeze/thawing releases the soluble mediators from the supernatants. (A) IL-6 levels as measured by Elisa in cell culture supernatants of Mewo cells that were treated with 0.2 μg/ml rWNT5A or carrier for 3 h and subsequently was freshly run on Elisa (4°C) or freeze/thawed for two cycles -80°C/4°C before analyses. The bar graphs show average fold increase as compared to carrier (ctrl) for each treatment indicated. Error bars represent SD. n = 2. (B) VEGF levels as measured by Elisa in cell culture supernatants of Mewo cells that were treated with 0.2 μg/ml rWNT5A or carrier for 3 h and subsequently was freshly run on Elisa (4°C) or freeze/thawed for two cycles -80°C/4°C before analyses. The bar graphs show average fold increase as compared to carrier (ctrl) for each treatment indicated. Error bars represent SD. n = 2. (C) MMP2 levels as measured by Elisa in cell culture supernatants of Mewo cells that were treated with 0.2 μg/ml rWNT5A or carrier for 3 h and subsequently was freshly run on Elisa (4°C) or freeze/thawed for two cycles -80°C/4°C before analyses. The bar graphs show average fold increase as compared to carrier (ctrl) for each treatment indicated. Error bars represent SD. n = 2. * = p <0.05; *** = p <0.001 by Student’s t-test.