Figure 6.
siRNA-mediated knockdown of MDK in three-dimensional (3D) paracrine assays. (A): Effect of MDK knockdown in SMCs on human umbilical vein endothelial cell (HUVEC) network formation in 3D paracrine coculture as per confocal microscopy over a 4-day timeline. (B): Quantification of the effect of MDK siRNA-mediated knockdown on total HUVEC network area in cocultures with the three SMC lineages on day 2. (C): Quantification of total cord length in cocultures containing MDK-siRNA-treated SMCs on day 2. (D): Quantification of the effect of MDK siRNA-mediated knockdown on the number of branch points in cocultures of the three SMC lineages on day 2 (∗∗, p < .01; ∗∗∗, p < .001; ∗∗∗∗, p < .0001; n = 3 independent biological replicates; scale bars = 100 μm). Abbreviations: D, day; LM, lateral mesoderm; MDK, midkine NE, neuroectoderm; ns, not significant; PM, paraxial mesoderm; siRNA, small interfering RNA; siScr, scrambled siRNA; SMC, smooth muscle cell.