Figure 5.

TLR3 signaling is required for efficient transdifferentiation of human fibroblasts to functional endothelial cells. (A) Fluorescent activated cell sorting (FACs) plot data obtained using CD144+ antibody to quantify iECs: (left panel) – scramble cells treated with vehicle control; (middle panel) – scramble cells treated with Poly I:C and (right panel) – TLR3-KD cells treated with Poly I:C. (B) Dot plot depicting FACs plot data obtained using CD144+ antibody to quantify iECs. All data represented as mean ± S.E.M, (n=3) *p<0.05, standard unpaired student t test, 1-way ANOVA corrected with Bonferroni method. (C) Representative images of iECs generated from Scramble treated cells (left panel) or TLR3-KD cells (right panel) showing reduced capacity to incorporate acetylated LDL by TLR3-KD cells. (D) Dot plot showing a significant decrease in the acetylated-LDL fluorescent intensity in TLR3-KD cells compared to scramble (n=3) *p<0.05, standard unpaired student t test and Mann-Whitney non-parametric test. (E) Representative images of iECs generated from scramble treated cells (left panel) or TLR3-KD cells (middle panel) showing failure to form capillary-like networks on matrigel by TLR3-KD cells. (F) Dot plot (right panel) showing a significant decrease in the number of nodes in TLR3-KD cells compared to Scramble (n=3) *p<0.05, standard unpaired student t test and Mann-Whitney non-parametric test. (G) Heat map of genes differentially expressed in iECs generated from scramble- or TLR3-KD fibroblasts showing 59 genes associated with angiogenesis reversed in the TLR3 KD-iECs when compared to scramble-treated iECs. (H) Dot plot depicting FACs plot data obtained using CD31+ antibody to quantify iECs: (left bar) – vehicle treated; (middle bar) control oligonucleotide treated with Poly I:C; (right bar) – P65 decoy (p65i) treated with Poly I:C.