HARE-Mediated Endocytosis of Hyaluronan and Heparin Is Targeted by Different Subsets of Three Endocytic Motifs

Figure 2

Hep binding to cell surface and total HARE in CD variants. Cells expressing human HARE (WT), the indicated HARE CD mutants, or EV were grown, washed, and preincubated in Endocytosis Medium at 37°C for 1 h to allow clearance of serum-derived glycosaminoglycans bound to HARE. Cells were chilled to 4°C, washed, and incubated with ¹²⁵I-SA•b-Hep complexes at 4°C and processed as described in Methods section to determine cell surface (a, c, e) or total cellular (b, d, f) specific ¹²⁵I-SA•b-Hep binding. Values are means ± SE (–9) and significant differences (assessed by Student’s -test) between WT and a HARE CD variant are indicated: ^#.