Fig 1: Characterization of the designed proteolytically resistant VEGF-B186R127S. (A) Graphical representation of VEGF-B186R127S. (B) Representative image of VEGF-B protein expression following HeLa cell transduction. (C) BaF3-VEGFR1 cell survival and proliferation assay. Each dot represents mean ± SD from two technical replicates from a representative experiment. (D) Receptor binding properties of different VEGF-B forms. Each lane represents the precipitation of ligands with hVEGFR1, hNRP1, hNRP2, or no receptor (blank). Red arrow signs indicate the full-length and proteolytically processed VEGF-B186, VEGF-B186R127S, or VEGF-A165.
Fig 2: VEGFR1 and VEGFR3 induce leptin production in trophoblasts.Western blot showing the effect of VEGFR1 siRNA (A) and VEGFR3 siRNA (B) on their respective receptor protein expression. Leptin secretion level measured in the media of BeWo cells treated with sFlt-1, siVEGFR1, siVEGFR3, siVEGFR1+ siVEGFR3, or a vehicle (C); Leptin secretion level measured in the media of BeWo cells treated with sFlt-1, VEGF165b, VEGFc, vehicle or a combination (D). Unpaired t-test in figure 3A–D. One Way ANOVA, Fisher’s LSD post hoc test for 3E-F. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001.
Fig 3: VEGFR1 and VEGFR3 are Predominant in Trophoblasts.Relative mRNA expression of VEGFR1, VEGFR2, VEGFR3 in both HUVECs and BeWo cells (A); VEGFR gene expression in BeWo cells (B); and HUVECs (C). BeWo cells treated with either a vehicle or sFlt-1 showing the protein levels of pVEGFR1 (D); and pVEGFR3 (E); One Way ANOVA, Tukey’s post hoc test for A-C. Unpaired t test in D,E. *p<0.05,***p<0.001, ****p<0.0001.
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