Figure 5: SARI inhibits angiogenesis through inhibiting Cp expression and the activity of the HIF-1α/VEGF axis. | Nature Communications

Figure 5: SARI inhibits angiogenesis through inhibiting Cp expression and the activity of the HIF-1α/VEGF axis.

From: RETRACTED ARTICLE: SARI inhibits angiogenesis and tumour growth of human colon cancer through directly targeting ceruloplasmin

Figure 5

(a) Immunoblots of SARI and HIF-1α expression in SW480 and HCT116 cells after stable expression of SARI. HCT116-control and HCT116-SARI cells were incubated under 1% O2 for 48 h or treated with DMOG for 24 h. β-Actin was used as a loading control. (b) Immunoblots of Cp, HIF-1α and VEGF expression in control and SARI cells with or without PX-478 (45 μM) or Tm (4.0 μM) treatment under normoxia and hypoxia. GAPDH was used as the loading control. (c) Immunoblots of Cp, HIF-1α and VEGF expression in control and SARI cells after transfection with or without p-Cp plasmid under normoxia and hypoxia. GAPDH was used as the loading control. (d) Expression of VEGF exacted by SW480-control and SW480-SARI cells with or without Tm (4.0 μM) and PX-478 (45 μM) treatment, after transfection with or without Cp expression plasmid (n=3; **P<0.01, NS, no significant difference; Student’s t-test). (e) Endothelial tube formation estimation following the incubation of HUVECs with conditioned medium from SW480-control and SW480-SARI cells with or without Tm treatment (4.0 μM) and PX-478 (45 μM), after transfection with or without Cp expression plasmid. The number of branches were quantified (n=5; ** P<0.01; NS, no significant difference; Student’s t-test). Scale bar, 50 μm. (f) Representative photomicrographs of HUVECs that have invaded through Matrigel chambers after incubation with conditioned medium from SW480-control and SW480-SARI cells with or without Tm treatment (4.0 μM) and PX-478 (45 μM), after transfection with or without Cp expression plasmid. Quantification of HUVECs that have invaded through Matrigel chambers (n=5; **P<0.01; NS, no significant difference; Student’s t-test). (g) Immunoblots of Cp and HIF-1α expression in HCT15, HCT15-shNC, HCT15-shSARI-1068 and HCT15-shSARI-554 cells with or without Tm (4.0 μM) treatment. GAPDH was used as a loading control. (h) Expression of VEGF exacted by HCT15, HCT15-shNC, HCT15-shSARI-1068 and HCT15-shSARI-554 cells with or without Tm treatment (4.0 μM) was measured by ELISA (n=3; **P<0.01; ##P<0.01; analysis of variance analysis).

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