Heparanase increases SDF-1 turnover in the BM.(A) SDF-1 levels in the BM of WT or hpa-Tg mice, detected by ELISA (i). Migration of cord blood CD34⁺ cells toward RPMI supplemented with supernatant from BM of WT or hpa-Tg mice placed in the lower chamber (ii). Data are means plus or minus SE; n ≥ 9 samples of BM supernatants. (B) mRNA levels of SDF-1 in BM cells from WT or hpa-Tg mice. Data are means plus or minus SE; n ≥ 4. (C) SDF-1 levels in the conditioned medium of MS-5 cells with or without pretreatment of the cells with heparanase, detected by ELISA (i). Migration of cord blood CD34⁺ cells toward the conditioned medium of MS-5 cells placed in the lower chamber of a transwell (ii). Data are means plus or minus SE; n ≥ 10 samples of conditioned medium. (D) (i) mRNA levels of SDF-1 (vs HPRT) in osteoblasts from WT or hpa-Tg mice. Data are means plus or minus SE; n ≥ 4. (ii) SDF-1 levels in the conditioned medium of osteoblasts obtained from WT or hpa-Tg mice, detected by ELISA. Data are mean plus or minus SE; n = 9. (E) Levels of biotinylated SDF-1 added exogenously to the BM supernatant of WT or hpa-Tg mice, detected by ELISA. Data are means plus or minus SE; n ≥ 8 samples of BM supernatants. (F) Levels of biotinylated SDF-1 added exogenously to the BM supernatant of hpa-Tg mice with or without pretreatment with a broad range protease inhibitor, detected by ELISA. Data are means plus or minus SE; n = 3 samples of BM supernatants (*P < .05).