Figure 7.

The effect of VEGFA and VEGFC on GAG charge. A: GEnCs incubated with vehicle, VEGFA, or VEGFC with [³H]glucosamine radiolabel to measure GAG metabolism and were separated into cell lysate and CM. Samples were loaded onto a DEAE ion exchange column; fractions were eluted in a NaCl gradient, collected, and read on a scintillation counter. Total disintegration counts are shown for CM and lysate. B and C: Counts are shown for low (0.4 to 0.5 mol/L NaCl), medium (0.5 to 0.6 mol/L NaCl), and high (0.6 to 0.7 mol/L NaCl) charged fractions in CM (B) and lysate (C). D: Percentage of low, medium, or high sulfated fractions in CM and lysate combined. mRNA was extracted from GEnCs treated with vehicle, VEGFA, or VEGFC for 1 hour. Real-time PCR analysis of NDST2 expression was quantified and normalized to GAPDH. Data are expressed as means ± SEM (A, B, C, and E). One-way analysis of variance (A and E, overall P < 0.01 and P < 0.05 respectively) or two-way analysis of variance (B and D, overall P < 0.01 and P < 0.05 respectively). n = 4. Bonfferoni post hoc tests indicated on graphs. ^∗P < 0.05 versus vehicle (E); ^∗∗P < 0.01 CM versus lysate (A), high versus low (B), or VEGFC versus vehicle (D).