Fig 1: UBE2O is the hub gene on saliva-Exos upregulated genes. a Valcano plot illustrating the differential mRNA expression level between saliva and saliva-Exos. The negative Log10 p values (y axis) are plotted against the Log2 fold changes in expression (x axis). b–e Analysis of biological process (BP) (b), cellular Component (CC) (c), molecular function (MF) (d) and KEGG pathway (e) of upregulated genes, the bubble diagram generated using the ggplot2 tool of the R software showing the top ten pathways. (f) Identification of UBE2O as one of the main hub genes using the DMNC algorithm to screen the upregulated genes. (g) UBE2O mRNA level in saliva and saliva-Exos assessed by qRT-PCR. n = 5. h, i UBE2O expression in HUVECs treated with PBS (control), saliva, and saliva-Exos, detected by Western blotting and qRT-PCR. ***p < 0.001, ****p < 0.0001
Fig 2: UBE2O mRNA is the one of the main cargo of Saliva-Exos, which could be transport into HUVECs, thereby decreases the level of SMAD6, subsequently activating BMP2, which, in turn, induces angiogenesis in vitro and promotes wound repair in vivo
Fig 3: UBE2O mediates HUVEC function by suppressing the SMAD6/BMP2 pathway. a Network map identifying UBE2O target genes. b Assessment of the expression of SMAD6 and BMP2 by Western blotting. c Expression of SMAD6 and BMP2 in HUVECs after indicated treatments evaluated by Western blotting. d, e Proliferation of HUVECs evaluated by the CCK8 and EdU assays. f Detection of the cell cycle-related proteins cyclin D1 and cyclin D3, by Western blotting. g Flow cytometry analysis of the cell cycle. h, i HUVEC migration evaluated by wound scratch and transwell assay. j Representative images of the tube formation by HUVECs after indicated treatments. k, l Quantitative analysis of total branch length and total number of branching points. *p < 0.05, **p < 0.01, ***p < 0.001
Fig 4: UBE2O promotes HUVEC function in vitro. a The UBE2O mRNA level was assessed by qRT-PCR after HUVECs were incubated with indicated treatments. b, c Proliferation of HUVECs after treatment with PBS, saliva, and saliva-Exos, measured by the CCK8 and EdU assays. d Distribution of HUVECs in the cell cycle assessed by flow cytometry. e–g Expression of cell cycle-related proteins determined by Western blotting. Evaluation of the migration of HUVECs by wound scratch assay and transwell assay. h Representative images of the tube formation assay in HUVECs treated with PBS, UBE2O, siUBE2O, and the combination of saliva-Exos and siUBE2O. i, j Quantitative analysis of total branch length and total number of branching points. *p < 0.05, **p < 0.01, ***p < 0.001
Fig 5: UBE2O promotes wound healing in vivo. a Representative images of wounds among two groups at indicated days. b A higher wound healing rate was seen in the UBE2O group than in the siUBE2O group. c H&E images of the wound sections in the two groups. d Smaller scar widths were seen in the UBE2O than in the siUBE2O group. e Blood flow at the wound sites assessed by laser speckle contrast imaging and quantification of the MPU ratio in the two groups. f Representative images of CD31-stained wound sections. g Quantification of the number of CD31-positive cells in the two groups. ****p < 0.0001; n = 6 per group
Supplier Page from Abcam for Anti-UBE2O antibody