The anti-angiogenic action of IGFBP-6 is IGF-independent. A) Tube formation assay was performed using HUVECs in the absence (panel a) or presence of mIGFBP-6, an IGFBP-6 mutant with impaired IGF binding (panel b: 500 ng/ml, and panel c: 1000 ng/ml). The effect was quantified by measuring the average tube length (B) and node number (C). Results, are expressed as a percentage of control (mean ± SE, n = 6. *, P<0.05; **, P<0.01; ***, P<0.0001 vs. control. ns, not significantly different).