Fig. 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 1μM SA, 10μM SA+ 0.3nM PTENi, PTENi (0.3nM), SA+ 100ng/mL VEGF-C, VEGF-C (100ng/mL), SA+ 100nM insulin or insulin (100nM) (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 10μM SA, media containing 10μM SA with PTENi (0.3nM), 10μM SA with VEGF-C (100ng/mL) or 10μM SA with insulin (100nM) for 12hrs. (Scale bar=25μm). 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).