Figure 2.

MM cell-derived Jagged promotes angiogenesis: (A) Tube formation assay on HPAECs with conditioned media (CM) of HMCLs^SCR or HCMLs^KD. 4X magnification images are shown on the left. Graphs (on the right) show the percentage variation of areas and branch points and total tube length +/-SEM. (B) Adhesion to fibronectin of HPAECs treated with CM from HMCLs^SCR and HMCLs^KD and stained with Calcein-AM. The graph reports the intensity of the adherent fluorescent cells. (C) Motility of the HPAECs treated with CM of HMCLs^SCR and HMCLs^KD was assessed by wound healing assays. Up: Representative pictures at 4X magnification. Down: The graph shows the average open area of the wounds expressed in pixels. (D,E) Variation of vascular endothelial growth factor (VEGF) expression in HMCLs^SCR and HMCLs^KD assessed at the mRNA level (D) by qRT-PCR of the relative gene expression variation (normalized to GAPDH) calculated by the 2^−ΔΔCt formula (data are expressed as the mean value ± SD) and at the protein levels (E) by ELISA on 48 h CM. Data are expressed as the amount of VEGF-A released by HMCLs^KD normalized on VEGF-A expressed by HMCLs^SCR. For each sample, the amount of VEGF-A (pg/mL) was normalized to the cell concentration. Statistical analyses were carried out by one-tailed t-tests; * is for p ≤ 0.05; ** is for p ≤ 0.01; *** is for p ≤ 0.001.