Fig 1: ADAMTS-1 downregulated the PI3K/Akt-eNOS-VEGF signalling pathway in HUVECs. HUVECs cultured with the supernatant from the pcDNA3.1-ADAMTS-1 transfection group or the pcDNA3.1 transfection group were incubated for 24 h with 740Y-P or A23187, which was followed by Western blot analysis. *P<0.05; **P<0.01. VEGF, vascular endothelial growth factor.
Fig 2: ADAMTS-1 inhibited the proliferation, migration and angiogenesis of HUVECs through the PI3K/Akt-eNOS-VEGF pathway. A549 cells were transfected with pcDNA3.1-ADAMTS-1 or pcDNA3.1. HUVECs were transfected with the pcDNA3.1-VEGF. (A) Effects of 740Y-P and VEGF overexpression on the proliferation of HUVECs cultured with supernatants from ADAMTS-1-overexpression A549 cells. Cell proliferation was examined by MTT assay; (B,C) influences of 740Y-P and VEGF overexpression on the migration of HUVECs cultured with supernatants from ADAMTS-1-overexpression A549 cells. Scratch assays were performed to measure cell migration; (D,E) regulation of HUVEC tube formation by 740Y-P and VEGF overexpression after being cultured with supernatants from ADAMTS-1-overexpression A549 cells. HUVEC tube formation was detected by angiogenesis assay. *P<0.05; **P<0.01. VEGF, vascular endothelial growth factor.
Fig 3: ADAMTS-1 inhibited in vitro angiogenesis of HUVECs. The supernatants of A549 cells with or without ADAMTS-1 overexpression were used to culture HUVECs. (A) Cell viability was analysed by MTT assay in HUVECs cultured with the supernatant from the pcDNA3.1-ADAMTS-1 transfection group or the pcDNA3.1 transfection group; (B) HUVEC cell-cycle distribution (S phase and G0/G1 phase) was analysed by flow cytometry; (C) scratch assays were performed to detect HUVEC cell migration; (D) tube formation of HUVECs was detected by angiogenesis assays. *P<0.05; **P<0.01.
Fig 4: Overexpression of ADAMTS-1 repressed VEGF expression and inhibited A549 cell proliferation. A549 cells were transfected with pcDNA3.1-ADAMTS-1 or pcDNA3.1. (A) The mRNA level of ADAMTS-1 in A549 cells was examined by qRT-PCR; (B) the protein level of ADAMTS-1 in A549 cells was examined by Western blotting; (C) relative VEGF mRNA levels in A549 cells with ADAMTS-1 overexpression detected by qRT-PCR; (D) the VEGF protein levels in A549 cells with ADAMTS-1 overexpression analysed by western blotting; (E) the VEGF protein levels in A549 cells with ADAMTS-1 overexpression detected by ELISA; (F) the viability of A549 cells with ADAMTS-1 overexpression by MTT assay. *P<0.05; **P<0.01. VEGF, vascular endothelial growth factor.
Fig 5: ADAMTS-1 was lowly expressed while VEGF was highly expressed decreased in A549 cells. (A) ADAMTS-1 and VEGF mRNA levels in A549 and BEAS-2B cells were examined by qRT-PCR; (B) ADAMTS-1 and VEGF protein abundances in A549 and BEAS-2B cells were determined by Western blotting; (C) ADAMTS-1 and VEGF contents in the culture supernatants of A549 and BEAS-2B cells by ELISA. *P<0.05; **P<0.01. VEGF, vascular endothelial growth factor; ELISA, enzyme linked immunosorbent assay.
Supplier Page from Abcam for Human ADAMTS1 ELISA Kit