Fig 1: Putative roles of YULINK in PDGF- or MCT-induced PAH pathological aberrations. During PAH-related pathogenesis induced by PDGF or MCT, an increase in YULINK expression occurs. YULINK coordinates with GLUT1 to facilitate glucose uptake, accompanied by an upregulation of HK-2, promoting enhanced glycolysis in PASMCs. This increased YULINK expression also contributes to uncontrolled cell migration and proliferation through the PDGFR-PI3K-AKT signaling pathways. While numerous reports have highlighted the role of this pathway in glucose metabolism and the Warburg effect, whether the triggered PI3K-AKT signaling regulates PAH-related processes requires further investigation. Suppression of YULINK through gene knockdown or the use of an inhibitor LY294002 to suppress PI3K activation has the potential to reverse these pathological abnormalities associated with PAH. In this graph, the black arrows represent the signaling transduction pathways, and the red arrows denote the enhancements in response to PAH pathogenesis. The dotted lines summarize findings from other published literature, not experiments conducted in this manuscript
Fig 2: Enhanced YULINK expression in MCT-induced PAH rats and human PAH specimen. A Pulmonary artery tissues were derived from normal and MCT-induced PAH rats for IHC staining. The brown color in the photomicrograph indicates the expression of YULINK expression. B Tissue sample from the right pulmonary artery of a clinical patient with severe PAH was subjected to IHC staining to assess YULINK expression. A normal pulmonary artery tissue was used as a control for comparison. Scale bar in 1–3: 100 µM, and 4: 200 µM
Fig 3: Positive correlation between YULINK expression and cell migration in PASMCs. YULINK expression in PASMCs with A YULINK knockdown (KD) or B overexpression (OE) was examined by western blot analysis. SC stands for scramble control. The numbers labeled below the respective blot lanes indicate the relative fold normalized with the internal control. C, D Micrographs (magnification 200×) together with bar graphs depict PDGF (5 and 20 ng/ml)-triggered cell migration in PASMCs with C YULINK KD or D YULINK OE using Transwell analysis. The values represent the mean of three independent experiments ± standard deviation. *p < 0.05 compared to scramble control without PDGF. #p < 0.05 between compared groups. E PASMCs with or without YULINK KD or OE were treated with PDGF in a dose-dependent manner (0, 5, and 20 ng/ml) for 24 h and subjected to western blot analysis for the indicated proteins. β-actin served as an internal control
Fig 4: Positive correlation between YULINK expression and cell proliferation in PASMCs. A PASMCs with YULINK KD or OE were treated with or without PDGF (20 ng/ml) for 5 days before harvest for MTT analysis. B PASMCs with the same treatment for 24 h were stained with PI and subjected to flow cytometry. The cell cycle profiles are presented with bar graphs indicating the distributions of the cells in the cell cycle. The values represent the mean of three independent experiments ± standard deviation. *p < 0.05 compared to scramble control without PDGF. #p < 0.05 between compared groups
Fig 5: YULINK knockdown reversed the enhancement of cell migration and proliferation in PAH-PASMCs. PASMCs and cells derived from MCT-induced PAH rats with or without YULINK KD were prepared for Transwell cell migration analysis. A Microscopy images presented with bar graphs indicate cell migration in PASMCs. Magnification ×200. B The cells were incubated with PDGF (20 ng/ml) for 24 h before harvest for protein extractions. Western blot analysis indicates the activation and expression of the indicated proteins. β-actin served as an internal control. The numbers labeled below the respective blot lanes indicate the relative fold normalized with the internal control. C MTT analysis of the 5-day proliferation of the cells. D The production of pyruvate in the cells. The values represent the mean of three independent experiments ± standard deviation. *p < 0.05 compared to normal PASMCs. #p < 0.05 between compared groups
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