Fig. 7. Upregulated PI3K/Akt and JAK/STAT3-mediated signalling are involved in sustaining vasculogenesis in a microfluidic system with an AdECM-containing gel. (a) Relative expression of differentially expressed genes in vasculature cultivated in a culture plate or a microfluidic device (Dynamic) with 0.5% AdECM-containing fibrin gel, as measured by real-time PCR (qPCR). (b) Immunofluorescence staining of the vascular network with pSTAT3 (green) and F-actin (red) after 14 days of cultivation in the presence or absence of static that has inhibitory effects against STAT3 to impede JAK/STAT3 pathway. (c and d) Vessel length and diameter were measured and quantified. (e) Immunofluorescence labeling of the vascular network was performed using pAkt3 (green) and F-actin (red) after cultivating the vasculature for 14 days with or without the addition of LY294002, an inhibitor of PI3K that blocks PI3K/Akt signaling. (f and g) The length and diameter of the vessels were assessed and quantified. Scale bar = 100 μm. *P < 0.05, compared to culture plate condition. ^#P < 0.05, compared to dynamic condition without inhibitor treatment, by Student's t-test. All data are presented as the mean (±SD) of three independent experiments.