Fig 1: EFNA3 inhibited tumor angiogenesis, cell invasion, and proliferation in PDAC cells. (a–b) The angiogenic abilities of the indicated treated HUVECs measured by tube formation assays. Magnification, ×200. (c–d) The invasion abilities of HUVECs were measured by transwell assays. Magnification, ×200. (e–f) The proliferation abilities of the indicated treated HUVECs were measured by EdU assays. Magnification, ×200.
Fig 2: miR-210 regulates EFNA3 expression. (a) The prediction for miR-210 binding sites on the EFNA3 transcript. (b) Luciferase activity in HUVECs cotransfected with the indicated miR-210 concentration or EFNA3 luciferase reporter transcript. Data are shown as the ratio of firefly activity to Renilla luciferase activity. (c) Relative EFNA3 expression in the indicated HUVECs was measured by qRT-PCR. (d) Relative EFNA3 protein expression in the indicated HUVECs was measured by WB analysis.
Fig 3: miR-210 or EFNA3 participates in the PI3K/AKT/VEGFA or Wnt/?-catenin/RHOA pathways. (a) Relative indicated protein expression in the indicated HUVECs was measured by WB analysis. (b) Dil-labeled exosomes were added to Calcein AM-labeled HUVECs for cocultivation. Images were captured with a fluorescence microscope, and the white arrows indicate exosomes. Magnification, ×100. (c) Dio-labeled exosomes of ov cy5-miR-210 Hs766T were added to HUVECs for cocultivation. Images were captured with a fluorescence microscope. The green arrow indicates exosomes, and the pink arrow indicates cy5-miR signals. Magnification, ×200.
Fig 4: Exosomal miR-210 promotes tumor progression via EFNA3 PI3K/AKT/VEGFA or EFNA3/Wnt/Β-catenin/RHOA (a) The in vitro angiogenesis abilities of the indicated treated HUVECs were measured by tube formation assays. Magnification, ×200. (b) Relative expression of the indicated proteins in the indicated HUVECs was measured by WB analysis. (c) In the transwell assay, the tumor cells that passed through the endothelial monolayer were imaged by fluorescence microscopy. Magnification, ×100.
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