Journal: BMC Pulmonary Medicine

Article Title: Astragaloside IV restrains pyroptosis and fibrotic development of pulmonary artery smooth muscle cells to ameliorate pulmonary artery hypertension through the PHD2/HIF1α signaling pathway

doi: 10.1186/s12890-023-02660-9

Figure Lengend Snippet: AS-IV reduced pyroptosis and fibrosis development in hypoxia-treated PASMCs. PASMCs were divided into 3 groups: Nor, Hyp and AS, cultured with normoxia or hypoxia, and treated with or without AS-IV. A - D , Cropped blots of GSDMD-N, NLRP3, cleaved Caspase-1, Fibronectin and Collagen1 and quantification of grayscale value (the samples derived from the same experiment and that gels/blots were processed in parallel). E , IL-1β and IL-18 levels in cell supernatant were measured by ELISA. F , Cell death was detected by PI/Hoechst double fluorescent staining. Scale bar = 50 μm. G, LDH activity was detected by a LDH release agent. H - I , Proteins related to extracellular matrix metabolism, including MMP2/9 and TIMP4 were detected by ELISA method. * P <0.05, ** P <0.01, *** P <0.001, comparison with Nor group. # P <0.05, ## P <0.01, ### P <0.001, comparison with Hyp group

Article Snippet: The nuclei were counterstained with Hoechst 33,258 (Abcam, #ab228550), and the image was visualized under a microscope and analyzed by the image J software.

Techniques: Cell Culture, Derivative Assay, Enzyme-linked Immunosorbent Assay, Staining, Activity Assay, Comparison

Journal: BMC Pulmonary Medicine

Article Title: Astragaloside IV restrains pyroptosis and fibrotic development of pulmonary artery smooth muscle cells to ameliorate pulmonary artery hypertension through the PHD2/HIF1α signaling pathway

doi: 10.1186/s12890-023-02660-9

Figure Lengend Snippet: AS-IV depressed pyroptosis and the production of fibrosis markers induced by hypoxia in PASMCs through the PHD2-HIF1α axis. A - D , PASMCs were divided into 5 groups: Nor, Hyp, AS, AS + si-PHD2 and LW6. Cropped blots of GSDMD-N, NLRP3, cleaved Caspase-1, Fibronectin and Collagen1 (the samples derived from the same experiment and that gels/blots were processed in parallel), and quantitation of the pyroptosis markers and the fibrosis markers. E , Cell death was detected by PI/Hoechst double fluorescent staining. Scale bar = 50 μm. F , IL-1β and IL-18 levels in cell supernatant were measured by ELISA. G, LDH activity was detected by a LDH release agent. H - I , Proteins of MMP2/9 and TIMP4 were detected by ELISA method. * P <0.05, ** P <0.01, *** P <0.001, comparison with Nor group. # P <0.05, ## P <0.01, ### P <0.001, comparison with Hyp group. ^ P <0.05, ^^ P <0.01, ^^^ P <0.001, comparison with AS group

Article Snippet: The nuclei were counterstained with Hoechst 33,258 (Abcam, #ab228550), and the image was visualized under a microscope and analyzed by the image J software.

Techniques: Derivative Assay, Quantitation Assay, Staining, Enzyme-linked Immunosorbent Assay, Activity Assay, Comparison

Journal: Molecular & Cellular Proteomics : MCP

Article Title: Decreased NAD Activates STAT3 and Integrin Pathways to Drive Epithelial-Mesenchymal Transition *

doi: 10.1074/mcp.RA118.000882

Figure Lengend Snippet: Lowering cellular NAD levels decreases cell growth rate. A, Western blotting confirmed CD38 overexpression in A549 cells. B, qPCR analysis confirmed overexpressed CD38 mRNA levels in A549 cells (n = 3). C, Western blotting confirmed CD38 overexpression in HepG2 cells. D, E, Cellular NAD and NADH levels in CD38(−) and CD38(+) A549 cells (n = 3). F, Growth curves of CD38(−) and CD38(+) A549 cells determined by CCK-8 assay (n = 3). G, Tumor samples harvested from mice 37 days after injection (n = 3). Tumor volumes (mm3) were measured using digital calipers and calculated using the formula: π/6 × length (mm) × width2 (mm). H, Growth curves of untreated- and 10 nm FK866-treated A549 cells determined by CCK-8 assay (n = 3). Data were analyzed using Student's t test. *p < 0.05; **p < 0.01; ***p < 0.001. *p < 0.05 was considered statistically significant. All values represent mean from at least three biological replicates ± S.E.

Article Snippet: After 36 h incubation, cells were treated with hydrogen peroxide (Aladdin, Shanghai, China) and cisplatin (Selleck, Houston, TX) in triplicate for 24 h. Cell counting kit-8 (CCK-8) reagent was added to treated cells and incubated at 37 °C for 2 h. Optical density (OD) was measured at 450 nm with a microplate reader (Bio-Rad).

Techniques: Western Blot, Over Expression, CCK-8 Assay, Injection