Fig 1: In vitro and in vivo experimental validation and GSEA analysis of TXNRD1. (A) The hemodynamic data and HE staining of lung tissue in animal experiments. (B) Representative Western blots and quantification of TXNRD1 and β-actin in the lungs of monocrotaline treated rats and controls (n = 7 each, *P < 0.05), data represent the mean ± SEM and Student t-test was used to compare two groups. (C) Immunofluorescence images of lung distal PA from MCT rats and controls. (D) Representative Western blots and quantification of TXNRD1 levels in rat PASMCs under PDGF-BB stimulation (30 ng/ml) normalized to a β-actin internal control (n = 3 each,*P < 0.05). (E) Venn diagram of GSEA terms among five datasets based on correlation analysis of TXNRD1. (F) The overlapping three GSEA terms correlated with TXNRD1. ****P < 0.0001.
Fig 2: Validation of TXNRD1 in clinical samples. (A) Gene expression of hub genes in GSE113439. (B) Serum TXNRD1 was decreased in IPAH patients. (C) The ability of serum TXNRD1 levels to diagnose IPAH was assessed using ROC curve analysis. (D,E) The relationship between serum TXNRD1 and mPAP as well as PVR analyzed by Spearman’s correlation analysis.
Fig 3: Silencing of TXNRD1 exacerbated PDGF-induced PASMC malignant phenotype. (A) Knockdown efficiency of TXNRD1 was verified by qPCR and Western Blot. (B) PASMCs were transfected with siRNA for 48 h before treatment with PDGF-BB (20 ng/ml) for another 24 h, and western blotting was used to detect proliferative and apoptotic markers in cell lysates. (C) Normalized quantification of cleaved-PARP, BAX/Bcl-2 and PCNA expression (n = 3 each, *P < 0.05). (D) Cell proliferative ability was determined by EdU assay in siTXNRD1-PASMC under PDGF stimulation. (E) Calculation of EdU-stained cells rate (n = 6 each, *P < 0.05). (F) Cell migration was measured by scratch assay in siTXNRD1-PASMC under PDGF stimulation. (G) Calculation of scratch closed percentage (n = 6 each, *P < 0.05). Data represent the mean ± SEM. Student t-test and one-way ANOVA were used to compare two and multiple groups. **P < 0.01, ***P < 0.001,****P < 0.0001.
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