polyclonal rabbit anti-human il-1ra  (Genzyme)

Journal: BMC Cancer

Article Title: Bladder cancer-derived interleukin-1 converts the vascular endothelium into a pro-inflammatory and pro-coagulatory surface

doi: 10.1186/s12885-020-07548-z

Figure Lengend Snippet: 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

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-human IL-1ra antibody (Sigma-Aldrich, St. Louis, MO, USA) 1:200 in DAKO Real antibody diluent (DAKO, Glostrup, Denmark).

Techniques: Activation Assay, Incubation, Produced, Translocation Assay, Fluorescence

Journal: BMC Cancer

Article Title: Bladder cancer-derived interleukin-1 converts the vascular endothelium into a pro-inflammatory and pro-coagulatory surface

doi: 10.1186/s12885-020-07548-z

Figure Lengend Snippet: Impact of UBC cell SN on the integrity of the endothelial cell layer. a HUVECs grown on gelatine coated ECIS electrodes were treated with starvation medium (control) or UBC cell SN. The impedance was measured continuously up to 12 h after treatment. T24 cell SN led to marked breakdown of the endothelial impedance. RT4 cell SN had a lower impact whereas RT112 cell SN, UROtsa cell SN or starvation medium (control) did not cause significant alterations after 12 h; n = 5; ** P ≤ 0.01. b HUVEC cells were incubated for 12 h with T24 cell SN (T24) or starvation medium (control) and then analysed by immunofluorescence staining of β-Actin (red) and CD31/PECAM-1 (green). In comparison to the control (left), treatment with T24 cell SN (right) induced the disintegration of adherent junctions (CD31/PECAM-1) indicating a weakening of the endothelial barrier. c Inhibition of cytokine signalling partially conserved endothelial barrier function. Antibodies against IL-6, CXCL-1 slightly mitigated impedance breakdown. In contrast, IL-1ra strongly attenuated tumour mediated endothelial dysfunction, whereas IL-8 neutralization had no impact; n = 4; ** P ≤ 0.01

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-human IL-1ra antibody (Sigma-Aldrich, St. Louis, MO, USA) 1:200 in DAKO Real antibody diluent (DAKO, Glostrup, Denmark).

Techniques: Incubation, Immunofluorescence, Staining, Inhibition, Neutralization

Journal: BMC Cancer

Article Title: Bladder cancer-derived interleukin-1 converts the vascular endothelium into a pro-inflammatory and pro-coagulatory surface

doi: 10.1186/s12885-020-07548-z

Figure Lengend Snippet: Impact of IL-1 on bladder cancer progression. a IL-1ra staining intensity in a cohort of 105 patients with UBC measured semiquantitatively by IHC. Controls (benign urothelium) showed a higher IL1-ra expression than in patients with cancer. No significant difference was found between low grade and high grade tumours. The staining intensity score was defined as follows: no staining = 1, low intensity = 2, moderate intensity = 3 and high intensity = 4. * P ≤ 0.05. n.s. = not significant. Representative IHC images are shown in b - c . Scale bar corresponds to 500 μm. b IHC very low staining intensity (score = 1). c Intermediate staining intensity (score = 3). d High staining intensity (score = 4). e High IL-1β mRNA levels were linked to adverse clinicopathological features high grade disease, muscle invasiveness and tumour progression. MIBC: muscle invasive bladder cancer, NMIBC: non-muscle invasive bladder cancer; * P ≤ 0.05

Article Snippet: Slides were incubated overnight at 4 °C with a rabbit polyclonal anti-human IL-1ra antibody (Sigma-Aldrich, St. Louis, MO, USA) 1:200 in DAKO Real antibody diluent (DAKO, Glostrup, Denmark).

Techniques: Staining, Expressing