Figure 3

Effects of Slit2N and VEGF-C on total VEGFR-3 levels and VEGFR-3 surface presentation/internalization in L-LECs. (A) Representative Western blot analysis of surface and total VEGFR-3 in L-LECs after incubation with 10 nM Slit2N for indicated times. GAPDH: loading control. (B) Quantitative analysis of surface VEGFR-3 in L-LECs after incubation with 10 nM Slit2N for indicated times. Band intensities from Figure 3A were determined by densitometry. Ratio of surface VEGFR-3/GAPDH in untreated L-LECs (“0” lanes) was set to “1” and values of other ratios were calculated vs. this control. (C) Quantitative analysis of total VEGFR-3 in L-LECs after incubation with 10 nM Slit2N for indicated times. Band intensities from Figure 3A were determined by densitometry. Ratio of total VEGFR-3/GAPDH in untreated L-LECs (“0” lanes) was set to “1” and values of other ratios were calculated vs. untreated control. (D) Representative Western blot analysis of membrane-bound VEGFR-3 (Surface VEGFR-3) and total VEGFR-3 in L-LECs after incubation with VEGF-C alone [100 ng/ml], and 10 nM Slit2N pretreatment + VEGF-C for indicated times. GAPDH: loading control. (E) Quantitative analysis of surface VEGFR-3 in L-LECs after incubation with VEGF-C alone [100 ng/ml] (Control), and 10 nM Slit2N pretreatment + VEGF-C for indicated times. Band intensities from Figure 3D were determined by densitometry. Ratio of surface VEGFR-3/GAPDH of untreated L-LECs was set to “1” and values of all other ratios were calculated vs. untreated control. (F) Quantitative analysis of total VEGFR-3 in L-LECs after incubation with VEGF-C alone [100 ng/ml] (Control), and 10 nM Slit2N pretreatment + VEGF-C for indicated times. Band intensities from Figure 3D were determined by densitometry. Ratio of total VEGFR-3/GAPDH of untreated L-LECs was set to “1” and values of other ratios were calculated vs. untreated control. Data for (B), (C), (E) and (F) represent the mean ± SD of 3 independent experiments (***p < 0.001).