Figure 6

Exposure of LECs to SA in vitro downregulates expression of VEGFR-3 and its downstream mediators. (a, b) Representative western blot images (a) for VEGFR-3, p-AKT, p-eNOS and quantification (b) normalized to GAPDH expression of total cellular protein harvested from LECs treated with 1 μm SA, 10 μm SA+0.3 nm PTENi, PTENi (0.3 nm), SA+100 ng ml⁻¹ VEGF-C, VEGF-C (100 ng ml⁻¹), SA+100 nm insulin or insulin (100 nm) (n=4–5/group, *P<0.05 for all antibodies). (c) Left panels: Representative immunofluorescent co-localization of VEGFR-3 (green) and Prox1, Ki67 and p-AKT (all shown in pink) in LECs cultured in control media, media containing 10 μm SA, media containing 10 μm SA with PTENi (0.3 nm), 10 μm SA with VEGF-C (100 ng ml⁻¹) or 10 μm SA with insulin (100 nm) for 12 h (scale bar=25 μm). Right panels: Quantification of expression of Prox1, Ki67 and p-AKT in groups shown in the left panels (n=10–15/group, *P<0.0005).