Fig. 3

CS-GAG modification is critical for SRGN-mediated cell migration. a Western blot analysis of SRGN in the total cell lysate and CM of H1299/Mock, H1299/SRGN and H1299/SRGN(S/A) cells cultured in serum-free medium for 48 h. b Wound healing assay was performed in H1299/Mock, H1299/SRGN and H1299/SRGN(S/A) cells. After 72 h, wound area (lower panel) was assessed by ImageJ and normalized to 0 h. c Boyden chamber migration assay was performed in H1299/Mock, H1299/SRGN and H1299/SRGN(S/A) cells. Migrated cells were counted in 3 h. d Boyden chamber migration assay was performed in HuTu80/CD44 cells that were suspended in CM derived from H1299/Mock, H1299/SRGN or H1299/SRGN(S/A) cells, respectively. Migrated cells were counted in 5 h. e CM harvested from H1299/Mock and H1299/SRGN cells were treated with or without ChaseABC for 24 h, and subjected to western blot analysis of SRGN. HuTu80/CD44 cells were suspended in non-treated or ChaseABC-treated CM and subjected to Boyden chamber migration assay. Migrated cells were counted in 5 h. Data are presented as the mean ± SD of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001 by Student’s t-test