Fig. 3

Pro-inflammatory activation of endothelial cells after stimulation with tumour cell SN. a After incubation with T24 cell SN, CXCL-1, IL-6, IL-8, GM-CSF and PAI-1 were significantly released from HUVECs (HUVECs + T24 cell SN). The SN of UROtsa was not able to stimulate HUVECs (HUVECs + UROtsa SN). HUVECs: baseline cytokine levels produced by HUVECs; T24 and UROTsa cell SN: baseline levels of cytokine produced by the tumour cells; (n = 8–12) ** P ≤ 0.01. b Incubation of HUVECs for 6 h with T24 cell SN promoted the trans-localization of NF-kB (red) into the nucleus (blue) of the endothelial cells. Treatment of HUVECs with the SN of RT112 and UROtsa cells had no effect. RT4 cell SN induced a weak trans-localization of NF-kB. Addition of IL-1ra blocked NF-kB translocation upon treatment with T24 cell SN. The bar diagram shows the fluorescence intensity of nuclear NF-kB. Scale bar corresponds to 50 μm; (n = 10 images) ** P ≤ 0.01. c IL-1ra inhibited the T24 cell SN induced release of the indicated pro-inflammatory cytokines from HUVECs. Cytokine levels were shown in pg/ml; (n = 3) * P ≤ 0.05