Journal: Advanced Science

Article Title: Homocysteine Exacerbates Pulmonary Fibrosis via Orchestrating Syntaxin 17 Homocysteinylation of Alveolar Type II Cells

doi: 10.1002/advs.202507803

Figure Lengend Snippet: Bulk‐seq and scRNA‐seq analysis of human IPF samples showing weakened Hcy metabolism in IPF. A) Schematic diagram of Hcy metabolism. B) GSEA showed the enrichment of fibrosis and Hcy metabolism terms in IPF group. C,D) Heatmap of enzymes ( MTR , MTRR , MTHFR , MTHFD1 , CBS , BHMT ) involved in Hcy metabolism and fibrosis markers ( COL1A1 , COL3A1 , ACTA2 , FN1 , CDH2 ). Data source: GSE52463 (C), GSE199949 (D). E) Correlation analysis between the expression of fibrosis and Hcy metabolism related genes in IPF environment. F) Spatial feature plots showing the distinct expression patterns of MTRR in control and IPF samples. G) T‐distributed stochastic neighbor embedding (t‐SNE) plot showing 13 distinct clusters resulting from scRNA‐seq of cells derived from lung tissues harvested from IPF and non‐IPF group. H) Joint density plot of Hcy metabolism genes created by scCustomize R package. I) Comparison of MTRR expression in all cells between control and IPF group demonstrated by t‐SNE plot. J) Violin plot showing the expression of MTRR among different cell types. K) Violin plot with dots showing the expression of MTRR in AT2 cells in control and IPF group. L) Pseudotime trajectory analysis of lung cells (AT1: alveolar type 1 cell; AT2: alveolar type 2 cell; Endo: endothelial cell; Basal: basal cell; Club: club cell; Fibro: fibroblast). M) MTRR relative expression with the progression of pseudotime in lung cells. N) Violin plot showing MTRR expression in different sub‐type cells of AT2 in control and IPF group (data source: GSE122960 , GSE190889 , GSE132771 ).

Article Snippet: After 1 h block at room temperature, membranes were reacted with the corresponding primary antibodies of which the details are Fibronectin (1:1000, T59537 , Abmart), Collagen I (1:1000, EPR7785, Abmart), Collagen III (1:1000, EPR17673 , Abmart), α‐SMA (1:1000, T55295 , Abmart, Shanghai), Vimentin (1:2000, ab92547, Abcam), N‐cadherin (1:10 000, ab76011, Abcam), CBS (1:5000, 14787‐1‐AP, Proteintech), MTRR (1:1000, 26944‐1‐AP, Proteintech), LC3B (1:1000, 14600‐1‐AP, Proteintech), ULK1 (1:1000, 20986‐1‐AP, Proteintech), SQSTM1/p62 (1:2000, ab109012, Abcam), Beclin‐1 (1:2000, T55092 , Abmart), STX17 (1:4000, 17815‐1‐AP, Proteintech), GAPDH (1:1000, T0004, Abmart), overnight at 4 °C.

Techniques: Expressing, Control, Derivative Assay, Comparison