Fig 8. LTBP2 reduces angiogenesis in vitro and in vivo.

(A) Expression of secreted LTBP2 in conditioned media from HONE1 and HK1 LTBP2-transduced cells. Coomassie blue staining of total protein in conditioned media was used to indicate equal loading. (B) LTBP2-conditioned media from HONE1 and HK1 cells suppressed the tube formation ability of HUVEC cultures. Bar graph represents an average of triplicate experiments ± S.E.M. (C) Matrigel plug tumors were stained for CD34 to visualize the microvessels, as indicated by the black arrows. LTBP2-transduced cells displayed significantly fewer microvessels in matrigel plugs compared to VA cells, as indicated by the bar graphs. (D) Real-time PCR of HONE1 (top) and HK1 (bottom) LTBP2- and VA-transduced cells showed the down-regulation of angiogenesis-related genes with LTBP2 expression: IL6, IL8, VEGF 165, VEGF 189, total VEGF, ANG, TSP1, uPAR, PDGB, RANTES, and MCP1. The housekeeping gene, GAPDH, was used as an internal control. The above data were obtained from three independent experiments in duplicate and the (*) for all graphs indicate P-value < 0.05.