Figure 7. Stimulation of matrix assembly varies with heparin chain length.

(A) CHO-677 cells were supplemented with FN alone (No Hep), or together with 2.8 μM unfractionated heparin (Hep), or with 11.2 μM of either H016 or H030 for the last 24 h of growth followed by immunostaining for FN and fluorescence microscopy to visualize the matrix. Scale bar indicates 50 μm. (B) DOC-insoluble matrix fractions of CHO-677 cells were isolated 24 h after addition of 25 μg/ml FN alone (0), or with the indicated μM concentrations of unfractionated heparin (Hep), H016, or H030. 2.8 μM heparin is equivalent to 50 μg/ml heparin. The mean fold change relative to unfractionated heparin is noted below. For three independent experiments, the average fold changes for H016 relative to Hep are 0.4 ± 0.1 for 2.8 μM, 0.4 ± 0.2 for 5.6 μM, and 0.6 ± 0.1 for 11.2 μM H016 (p < 0.05). For H030, average fold changes are 1.2 ± 0.2 for 2.8 μM, and 1.1 ± 0.4 for 5.6 and 11.2 μM H030. Representative blots are shown. For the right-hand blot, intervening lanes within the same gel were cropped.