Fig. 1

Glycosaminoglycan-modified betaglycan (BG) sheds more than unmodified BG. A, B Autoradiograph of samples after [¹²⁵ I]-TGF-β1 binding and crosslinking of either A total cell lysates or B from conditioned media of indicated cells with stable (HEYA8, SKOV-3) or transient (OVCA-429) expression of BG (control-vector, FL-BG, ∆GAG-BG) and C quantification of the cell surface binding autoradiograph to the conditioned media binding autoradiograph of indicated cells expressing FL-BG or ∆GAG-BG. D-G BG-ELISA from the conditioned media of indicated HEYA8 D, E and SKOV-3 F, G cells. The concentration of shed-BG normalized to the total protein concentration of the cells is plotted by Mean ± SEM, (n = 4 for stable and n = 3 for transient). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001, One-way ANOVA followed by unpaired t-test between FL-BG and ∆GAG-BG (HEYA8 stable, p = 0.0007, transient, p = 0.0018, FL-BG vs ∆GAG-BG) (SKOV-3 stable, p = 0.034, transient, p < 0.0001, FL-BG vs ∆GAG-BG). H Autoradiograph of SKOV-3 parental and BG CRISPR Knockout (BGKO) cells radiolabeled with [¹²⁵ I]-TGF-β1 followed by immunoprecipitation using anti-BG antibody. I Western blot of SKOV-3 BGKO cells transiently expressing FL-BG (BGKO-FL) or ∆GAG-BG (BGKO-∆GAG) immunoblotted with anti-BG. J BG ELISA of conditioned media collected from the SKOV-3 BGKO cells expressing FL-BG (BGKO-FL) and ∆GAG-BG (BGKO-∆GAG). The concentration of shed-BG normalized to the total protein concentration of the cells is plotted (Mean ± SEM, n = 5). ****p < 0.0001, One-way ANOVA followed by unpaired t-test between BGKO-FL and BGKO-∆GAG