Effect of the IL‐10—IL‐10R pathway on the angiogenic activity of HUVECs in vitro. (A, B) HUVECs were treated with medium alone (CON), rhIL‐10 (10 U/ml, unless otherwise indicated), hIL‐10 mAb (10 µg/ml, unless otherwise indicated), or hIL‐10R mAb (10 µg/ml, unless otherwise indicated) for migration assays and tube formation assays. (C, D) CM from treated ectopic EN‐MSCs (Ec) was cultured with HUVECs for the migration assay and the tube formation assay. CM from human eutopic EN‐MSCs (Eu) was used as a cell type control. HUVECs treated with medium alone served as a negative control (CON). (E) Representative western blotting for VEGF, p‐STAT3, total STAT3, and β‐actin in treated HUVEC cells as shown in A. (F) HUVECs were transfected with scrambled siRNA (scramble) or with VEGF‐specific siRNA at 25 or 50 nm for 12 h, followed by western blotting for VEGF and β‐actin in treated HUVECs. Each immunoblot was quantified for the target protein relative to its corresponding β‐actin levels. Results are shown as ratios that were normalised to the corresponding CON sample. These are representative blots from three independent experiments. (G, H) Tube formation of HUVECs after treatment with scrambled siRNA, VEGF siRNA at 25 nm in PBS, or VEGF siRNA at 25 nm together with rhIL‐10 for 12 h. Results (mean ± SD) shown in A–D and H are from three independent experiments.
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