Figure 4

A: DEAE-sepharose binding property of endocan and endocanΔ2. The crude supernatants (Input) of transiently transfected HEK 293 cells with endocan (grey box) or endocanΔ2 (white box) (n = 4). The bound materials were eluted in 1 M NaCl (Eluate). Endocan and endocanΔ2 levels were measured by ELISA (median values with error bars showing the range). B: The eluates from DEAE-Sepharose column were digested overnight with chondroitinase (Ch) ABC. Digested (lanes 2, 4, 6, 8) and undigested (lanes 1, 3, 5, 7) samples were analyzed under reducing (lanes 1–4) or nonreducing (lanes 5–8) conditions. Black arrow indicates the endocan translation product and white arrow the endocanΔ2 translation product. Stars indicate the multimeric forms of endocanΔ2. C: Western blot analysis of endocan immunoprecipitated from endothelial or transfected cells supernatants. Lane 1: HUVEC; lane 2: endocan-HEK 293; lane 3: endocanΔ2-HEK 293. Black arrow indicates the endocan and endocanΔ2 translation products. Star indicate the multimeric form of endocanΔ2; Arrowheads indicate the non-specific bands originating from the secondary Ab. D. Endocan (grey box) and endocanΔ2 (white box) levels in the supernatant and cell lysate of HEK293 cells after 72 hours of culture assessed by ELISA (median, with error bars showing the range, n = 6) **, p = 0.022.