Fig. 4

Unmodified BG (∆GAG) promotes invasion in a TGF-β signaling-dependent manner. A Representative images of invasion through Matrigel-coated Boyden-transwell inserts of HEYA8 cells expressing control vector, FL-BG or ∆GAG-BG, scale bar = 275 μm, and quantification of invaded cells. B Representative images of invasion through Matrigel-coated Boyden-transwell inserts of SKOV-3 BGKO cells as indicated and quantification of invaded BGKO cells. Mean ± SEM are plotted, (HEYA8 Stable n = 16, SKOV-3 BGKO, BGKO-FL, and BGKO-∆GAG, n = 8) ***p < 0.001; ****p < 0.0001, One-way ANOVA followed by unpaired t-test between FL-BG and ∆GAG-BG. (p < 0.0001, HEYA8 Stable FL-BG vs ∆GAG-BG, p < 0.0001, SKOV-3 BGKO-FL vs. BGKO-∆GAG). (C) Representative images of invasion through Matrigel-coated Boyden-transwell inserts of HEYA8 FL-BG and ∆GAG-BG cells treated with soluble recombinant BG at 1 ng/mL in the top chamber. Scale Bar = 275 μm and quantification of the invaded. Mean ± SEM are plotted, (n = 5), ****p < 0.0001, One-way ANOVA followed by unpaired t-test between sBG untreated to sBG treated groups. (p < 0.0001 ∆GAG-BG untreated vs. ∆GAG-BG, sBG treated). (D) Representative images of invasion through Matrigel-coated Boyden-transwell inserts of HEYA8 FL-BG and ∆GAG-BG cells treated with 500 nM of A83-01 in the top chamber and quantification of invaded cells. Mean ± SEM were plotted, (n = 4), ****p < 0.0001, One-way ANOVA followed by unpaired t-test between untreated compared to A83-01 treated groups. (p < 0.0001 ∆GAG-BG untreated vs. ∆GAG-BG, A83-01 treated)