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. 2021 Oct 8;11:753244. doi: 10.3389/fonc.2021.753244

Figure 5.

Figure 5

Inhibition of SHP-2 impairs AM-induced decreases of HUVECs permeability and AM ability to form capillary-like structures in vitro. (A) Paracellular permeability for 250 KDa FITC-dextran was determined for HUVECs, treated either with AM (10-9 M), or AM (10-9 M) in presence of NSC-87877 (30 μM), or SHP099 (30 μM), and cultured on Transwell filters. Permeability of control cells was set to 100% (n = 6 for each group). **, p < 0.01. (B) HUVECs were transfected with control siRNA (siCtrl) or with siRNAs directed against SHP-2 and cultured on Transwell filters for 24 h. Paracellular permeability for 250 kD FITC-dextran was determined. Permeability of control-transfected cells was set to 100% (n = 4 for each group). (C) The morphogenic activity of AM is impaired upon inhibition of SHP-2. HUVECs (7 x 104 cells/well) were seeded into Matrigel-precoated wells and cultured in low-serum conditions (0.5% FBS) in absence (Ctrl) or presence of AM (10-9 M), AM (10-9 M) + NSC-87877 (30 μM), AM (10-9 M) + SHP099 (30 μM). (D) Photographs were taken to cover the whole surface after treatment for 5 h, and capillary-like structures were quantified. Scale bar, 100 μm; *p < 0.05; **p < 0.01; ***p < 0.001. The value of control cells was set to 100%. Each experiment is representative of three (A, B), and four (D) independent experiments. Results are shown as means ± SD.