Figure 3: NGD16 inhibited pro-angiogenic signaling molecules. (A) PC-3 and MiaPaca-2 cells were left untreated or treated with NGD16 for 24 h. Whole cell lysates were checked for the expression of GRP78, VEGFR-2, MMP-9 and TIMP-1 through western blot analysis. (B) Expression of above angiogenic regulators were also observed through western blot analysis in endothelial HUVEC cells. (C) The ability of ectopic GRP78 to induce pro-angiogenic mediators like; MMP-9 and VEGFR-2 was examined in PC-3 and MiaPaca-2 cells in presence and absence of NGD16, 24 h posttransfection with plasmid construct, pCMV BiP-Myc-KDEL-wt/vector. (D) PC-3 and MiaPaca-2 cells were treated with DMSO or different concentrations of NGD16 for 48 h. Conditioned media was analyzed for MMP-9 gelatinase activity. The values in between the western blot and zymography bands indicate densitometry analysis of fold increase/ decrease of expression in digital format. Western blot and gelatin zymography results were subjected to statistical analysis for significance (n = 3, * p< 0.05).