The effect of VEGFA and VEGFC on uncharged GAGs. A and B: GEnCs incubated with vehicle, VEGFA, or VEGFC [3H]glucosamine radiolabel were separated into CM and lysate. 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 (A) and lysate (B), demonstrating equal experimental input. C and D: Total counts are shown for the fraction eluted in 0.3 mol/L NaCl (minimal charge) (C) and separated into CM and lysate (D). E: Uncharged ion exchange fractions were either digested in hyaluronidase (low molecular weight HA) or left undigested (high molecular weight HA) and injected onto a size-exclusion column. Dashed line divides LMW from HMW. Data are expressed as means ± SEM. n = 4. Data were analyzed by one way ANOVA (C, overall P < 0.05) or two way ANOVA (D, overall P < 0.05). Bonfferoni post hoc tests are indicated on graphs. ∗∗P < 0.01 versus vehicle (C), versus CM vehicle (D). MW, molecular weight.