Fig. 5.

Addition of Cyr61 restores the angiogenic activity of Cyr61-depleted MSC secretome. Immunodepletion was performed as described in Figure 4 and Experimental Procedures Section. A: After incubation, the secretomes were immunoblotted with anti-Cyr61. In all cases, 66 μg proteins per well were loaded. B: Morphogenesis of HPAE cells in matrigel induced by secretome, secretome depleted with anti-Cyr61 or normal IgG, and Cyr61-depleted secretome supplemented with 0.1 μg/ml human recombinant Cyr61 polypeptides (Cell Sciences). DMEM and 10% (v/v) FBS were used as negative and positive controls, respectively. C: Quantitative analysis of tubule length. Note that depletion of Cyr61 from MSC secretome completely abrogates the angiogenesis-inducing capability of MSC secretome. Also note that the addition of recombinant human Cyr61 restores the angiogenesis-promoting activity of the Cyr61-depleted secretome. The total tubule length quantification was done by addition of the tubule length from five random fields per well. Data are Mean ± SD of triplicate determinations. Scale bar, 400 μm.