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: Annals of Translational Medicine

Article Title: The landscape of m6A regulators in esophageal cancer: molecular characteristics, immuno-oncology features, and clinical relevance

doi: 10.21037/atm-22-5895

Figure Lengend Snippet: In vivo and in vitro functional assay involving VIRMA . (A) Western blot analysis of VIRMA expression efficiency in siRNA knockdown KYSE410 and OE19 cells. (B) The CCK-8 assay was used to detect the viability of KYSE410 and OE19 cells transfected with VIRMA siRNAs or control. (C) Western blot analysis of VIRMA expression efficiency in shRNA knockdown KYSE410 and OE19 cells. (D) The Transwell assay was performed to detect the migration and invasion ability of KYSE410 and OE19 cells transfected with VIRMA siRNAs or control. Stained with 3% crystal violet in methanol. (E) Subcutaneous tumorigenicity assay of KYSE410 and OE19 cells transfected with VIRMA shRNA or control, the right side is the H&E staining of the tumor. Data are representative of at least three independent experiments. Data are presented as mean ± SEM. ****, P<0.0001. NC, negative control; H&E, hematoxylin and eosin; SEM, standard error of the mean.

Article Snippet: Each day, one 96-well plate was used, and 10 µL/well of Cell Counting Kit-8 (CCK-8) (Bioss, Beijing, China) solution was added.

Techniques: In Vivo, In Vitro, Functional Assay, Western Blot, Expressing, CCK-8 Assay, Transfection, shRNA, Transwell Assay, Migration, Staining, Tumorigenicity Assay, Negative Control