Journal: Engineering

Article Title: PAI-1 Derived from Alveolar Type 2 Cells Drives Aging-Associated Pulmonary Fibrosis

doi: 10.1016/j.eng.2024.08.014

Figure Lengend Snippet: Fig. 4. Impact of the secretome from senescent epithelial cells on fibrosis and Collagen I synthesis. (a) Schematic of Collagen I biosynthesis; (b) schematic diagram of the cell culture models used for conditioned medium culture and coculture experiments in this study; (c) Western blot analysis of senescence- and cell cycle-associated proteins in primary AT2 cells treated with 30 gL1 D-galactose for 48 h (n = 3); (d–e) Western blot analysis of fibrosis, collagen synthesis, and degradation-related proteins in NIH3T3 cells cultured with conditioned medium from D-galactose-induced senescent (d) primary AT2 cells and (e) A549 cells for 48 h (n = 3). MMP1: matrix metalloproteinase.

Article Snippet: After 8 h of transfection, the NIH3T3 cells were exposed to medium containing 2 lg mL 1 PAI-1 (HY-P71133, MCE) and 0.4% serum.

Techniques: Cell Culture, Western Blot

Journal: Engineering

Article Title: PAI-1 Derived from Alveolar Type 2 Cells Drives Aging-Associated Pulmonary Fibrosis

doi: 10.1016/j.eng.2024.08.014

Figure Lengend Snippet: Fig. 5. PAI-1 secreted by senescent epithelial cells promotes collagen I synthesis via the TGF-b/Smad2/3 pathway. (a) Schematic diagram of the process used to screen for key SASP factors associated with fibrosis; (b) RT-qPCR analysis of key fibrosis-related SASP factors in 3 and 28 month old mouse lung tissues. Each group comprised six biological replicates and three technical replicates. Results are presented as mean ± standard deviation (SD); (c) expression analysis of key fibrosis-related SASP factors in primary AT2 cells from 3 and 24 month old mice (n = 3); (d) Western blot analysis of PAI-1 protein in mouse lung tissues of different ages and D-galactose-induced senescent A549 cells (n = 3); (e, f) Western blot analysis of fibrosis and collagen synthesis-related proteins in (e) NIH3T3 cells and (f) WI-38 cells stimulated with gradient concentrations of PAI-1 (0, 0.5, 1.0, and 2.0 lgmL1) for 48 h (n = 3); (g, h) activation of the TGF-b pathway in (g) NIH3T3 and (h) WI-38 cells after stimulation with 2.0 lgmL1 PAI-1 for 48 h (n = 3); (i) effect of PAI-1 on Col1a1 synthesis after TGF-b pathway inhibition with SB431542 (n = 3); (j) impact of PAI-1 on Col1a1 synthesis after silencing Smad2/3 with siSmad2/3 (n = 3).

Article Snippet: After 8 h of transfection, the NIH3T3 cells were exposed to medium containing 2 lg mL 1 PAI-1 (HY-P71133, MCE) and 0.4% serum.

Techniques: Quantitative RT-PCR, Standard Deviation, Expressing, Western Blot, Activation Assay, Inhibition