Fig 2: RDN improves atheroprone endothelial phenotypes and inflammatory response in HFD-fed ApoE-/- mice.Aortic root cross-sections were conducted co-immunofluorescent staining with anti-CD31 (green) and MCP-1 (red in A), ET-1 (red in B), ICAM-1 (red in C), or VCAM-1 (red in D) antibodies (n = 6). The MFI of the indicated molecules in vascular endothelium were quantitatively analyzed. E-G: Expression of TNF-a, IL-1ß and IL-6 in aortic root cross-section were determined by immunofluorescent staining with quantification of MFI (n = 6). H: Serum IL-6, IL-1ß, TNF-a, CXCL-10 and IL-10 levels were determined by the corresponding ELISA assay kits (n = 6). I: Aortic root cross-sections were conducted immunofluorescent staining with anti–NF–?B p65 antibody with quantitative analysis of MFI in the nucleus (n = 6). J: I?Ba protein expression in mouse thoracic aorta was determined by Western blot (n = 5). Data are presented as mean ± SD. *P < 0.05; **P < 0.01; ***P < 0.001. ns: not significant. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Fig 3: Immunostaining analysis of transplanted M-MSCs. Confocal micrographs of M-MSCs stably expressing GFP (red) and immunostaining of the marker proteins E-cadherin (Ecad), vimentin, a-SMA, and CD31 (green) in bladder sections of LPS-IC + M-MSC rats at 7 DAT (A) and 30 DAT (B) (magnification ×1,000, scale bar = 10 µm). Nuclei were stained with DAPI (blue). Arrows indicate GFP+ engrafted cells co-expressing each tissue marker. Notably, the majority of transplanted GFP+ cells at 7 DAT was strongly stained with the vimentin expressing stromal cells (second row in A) and perivascular cells (third row in A).

Fig 4: Morphological features of cultured cells obtained from CD, CP and NL tissue.A) Immunofluorescent staining in CD- and CP-Hep for Heppar1, CD31 (endothelial marker) and CD68 (macrophages marker) at 8 weeks. B) Representative immunohistochemical staining of Heppar1 in NL, CD, CP and HCC tissue utilized for cell isolation. Heppar1 showed overexpression in HCC compared to NL, CD and CP. C) Morphological features were observed at each time intervals (T1, T2, T3 and T4) and bright fields are captured (image magnification x100; black bar, 200µm).

Fig 5: LINC00961 knockdown attenuates injury and EndMT in HCMECs by activating the PTEN-PI3K-AKT signalling pathway. (A) Immunofluorescence staining was used to identify CD31 + /a-SMA+ cells. a-SMA (red), CD31 (green); scale bars: 50 µm. (B) Rate of cell apoptosis was evaluated using flow cytometry. (C) Reverse transcription-quantitative PCR was used to determine the expression of PTEN, PI3K, AKT and mTOR mRNA. (D) Western blotting was used to determine the protein expression of PTEN, p-PI3K, PI3K, p-AKT, AKT, p-mTOR and mTOR. (a) TGF-ß + sh-LINC00961, (b) TGF-ß, (c) TGF-ß + LV-LINC00961 and (d) TGF-ß + sh-LINC00961 + VOT. Each experiment was repeated at least three times. ns, P>0.05; *P<0.05, #P<0.05 and &P<0.05. n=5 per group. EndMT, endothelial-mesenchymal transition; HCMECs, human cardiac microvascular endothelial cells; PTEN, phosphatase and tensin homolog deleted on chromosome 10; a-SMA, a-smooth muscle actin; p-, phosphorylated; sh-, short hairpin; LV, lentivirus; VOT, VO-OHpic trihydrate; ns, not significant.