Fig. 3

circ-IARS increases the permeability of endothelial monolayer cells by up-regulating RhoA activity. a qRT-PCR was used to selected the better interference siRNA, siRNA 2. HUVECs transfected with circ-IARS overexpression plasmids were named the ov-circ-IARS group; HUVECs transfected with circ-IARS siRNAs were named the si-circ-IARS group. b-d Relative expression levels of circ-IARS (b), RhoA (c) and ZO-1(d) were measured by qRT-PCR. e Relative RhoA activity was significantly increased in the ov-circ-IARS group and decreased in the si-circ-IARS group. f Protein levels of RhoA, ZO-1, RhoA-GTP and F-actin were evaluated by western blotting. g HUVEC coverslip culture results revealed expression of F-actin and focal adhesion. Scale bars = 100 μm. h In a Transwell assay, tumor cells attached to the lower side of the filter were imaged (Olympus) and counted. Scale bars = 50 μm. i-k Hs 766 T cells, with circ-IARS up-regulation or NC, were injected into the head of the pancreas in animal experiments to establish a pancreatic cancer tumor model. Images were captured with IVIS Imaging System each week (i); a month later, the mice were sacrificed, and the liver or pancreas were checked (j). Furthermore, pancreatic cancer in situ or liver metastasis was confirmed by H&E staining; the blue arrows indicate pancreatic cancer foci, and the black arrows indicate metastatic liver foci. Scale bars = 50 μm (k)