Journal: Tissue Engineering Part A

Article Title: MicroRNA-1 and MicroRNA-206 Improve Differentiation Potential of Human Satellite Cells: A Novel Approach for Tissue Engineering of Skeletal Muscle

doi: 10.1089/ten.tea.2011.0191

Figure Lengend Snippet: FIG. 1. Satellite cell culture, differentiation, and characterization. During passages 8–15, cultured cells display a homoge- neous morphology, note the triangular-shaped cells that are typical for satellite cells, by differential interference contrast (DIC) microscopy (A). An immunofluorescent image of proliferating cells at passage 9 double stained for a satellite cell marker desmin (green) and a fibroblast marker MCA1399G (red). Nuclei are counterstained with DAPI (blue). More than 95% of the cells expressed the satellite cell marker desmin (B). Differentiation of confluent satellite cell cultures was induced by switching to differentiation medium. Five days after switching to differentiation medium, myotubes were clearly visible by DIC microscopy (C). All scale bars are 100 mm. Color images available online at www.liebertonline.com/tea

Article Snippet: The primary antibody consisted of either (1) a myogenic marker, rabbit-anti-human desmin (1:100; Novus Biological), (2) a fibroblast marker, mouse anti-human MCA1399G (1:100; AbD Serotec), (3) a sarcomere component, mouse-anti-human a-sarcomeric actin IgM (1:200; clone Alpha Sr-1; Abcam) (4) a myogenic transcription factor, mouse-anti-human MyoD (1:100; Dako), (5) a sarcomere component, mouse-anti-human myosin (MF20; 1:500), and (6) a satellite cell marker, mouse-anti-human Pax7 (1:10; both Developmental Studies Hybridoma Bank).

Techniques: Cell Culture, Microscopy, Staining, Marker

Journal: Nature Communications

Article Title: Targeted immunotherapy rescues pulmonary fibrosis by reducing activated fibroblasts and regulating alveolar cell profile

doi: 10.1038/s41467-025-59093-7

Figure Lengend Snippet: A Schematic diagram of fibrosis model establishment and treatment strategy in young or aged mice. B , C Young mouse BW changes ( B ) and survival curve ( C ) of control and BLM groups treated with IgG/ β LNP-FAPCAR 2.2, CD5/ β LNP-FAPCAR 2.2 (10 μg) and saline ( n = 10). The survival rate was analyzed using the log-rank (Mantel-Cox) test. D Lung coefficients (lung weight (mg)/BW (g)) ( n = 5,5,10,10,10,7). E tdTomato reporter gene expressed in T cells of spleen and lung at 24 h after the second CD5/ β LNP-FAPCAR 2.2 injection (Scar bar: 10 μm). F Flow cytometry plots for FAPCAR expression in CD3 + pulmonary T cells of BLM+Saline and CD5/ β LNP-FAPCAR 2.2 treatment groups and the ratio of FAPCAR expression to CD3 + pulmonary T cells ( n = 4). G Flow cytometry analysis of FAP + Pdgfra + fibroblasts in BLM+Saline and CD5/ β LNP-FAPCAR 2.2 treatment groups, ratio of FAP + Pdgfra + fibroblasts to all lung cells, and ratio of Pdgfra + fibroblasts (both FAP + and FAP — ) to all lung cells ( n = 4). H , I Representative micro-CT images and mean HU density ( H ), and lung air volume (mm 3 ) ( I ) computed according to CT results at postoperative 28 days ( n = 4,4,7,7,7,7). J Digital photos of lung tissue. ‘n’ means independent biological replications. The values are the means ± SDs. * P < 0.05. P -value determined by one-way ANOVA followed by Tukey’s multiple comparisons test ( D , F , H , I ) and two tailed unpaired Student’s t -test ( G ). (BW: body weight; HU: Hounsfield unit). Source data are provided as a Source Data file. Created in BioRender ( A , B ) .

Article Snippet: FVS 700 (live/death dye, #564997, BD Bioscience, CA, USA), CD3e (#562600, BD Bioscience), CD8a (#557654, BD Bioscience), F4/80 (#565411, BD Bioscience), CD86 (#558703, BD Bioscience), CD163 (#12-1631-82, Thermo Fisher), FAP (#FAB9727G, R&D system) and Pdgfra (#12-1401-81, Thermo Fisher) were used in experiments.

Techniques: Control, Saline, Injection, Flow Cytometry, Expressing, Micro-CT, Two Tailed Test

Journal: Redox Biology

Article Title: Lipid peroxidation triggered by the degradation of xCT contributes to gasdermin D-mediated pyroptosis in COPD

doi: 10.1016/j.redox.2024.103388

Figure Lengend Snippet: The primers.

Article Snippet: Primary antibodies against caspase-4/11, FSP1, GCH1 and GPx4 were obtained from Zenbio (Chengdu, China).

Techniques:

Journal: Redox Biology

Article Title: Lipid peroxidation triggered by the degradation of xCT contributes to gasdermin D-mediated pyroptosis in COPD

doi: 10.1016/j.redox.2024.103388

Figure Lengend Snippet: CSE triggered lipid peroxidation and pyroptosis in Beas-2b cells. (A) The LDH release rate was measured by the LDH cytotoxicity detection kits. (B) The GSH/GSSG ratio in treated cells were measured with the corresponding kit. (C) The levels of MDA in treated cells were measured with the corresponding kit. (D–F) ELISAs showed that CSE increased the levels of IL-1α, IL-1β and IL-18 in culture medium in a dose-dependent manner. (G) Lipid peroxidation in cells was examined by Liperfluo staining and observed by confocal microscopy (400 × ); ROS production was examined with a DCFH-DA probe and observed with a fluorescence microscope (200 × ). (H) The expression levels of xCT, GPx4, NLRP3, cleaved caspase-1/4, ASC, and cleaved IL-1β were detected by western blotting and analyzed by ImageJ. (I) The expression levels of FSP1 and GCH1 were tested by western blotting and analyzed by Image J. (J) The number of living cells and dead cells were detected by Calcein/PI staining (200 × ). (K) TEM showed the pyroptotic body formation, cell membrane and mitochondria. Green arrows indicate normal mitochondria, blue arrows indicate normal cell membranes, red arrows indicate abnormal mitochondria, and white arrows indicate damaged cell membranes (0.5 μm). NS p＞0.05, ∗p＜0.05,∗∗p＜0.01 compared with the control group.

Article Snippet: Primary antibodies against caspase-4/11, FSP1, GCH1 and GPx4 were obtained from Zenbio (Chengdu, China).

Techniques: Staining, Confocal Microscopy, Fluorescence, Microscopy, Expressing, Western Blot, Membrane, Control

Journal: Redox Biology

Article Title: Lipid peroxidation triggered by the degradation of xCT contributes to gasdermin D-mediated pyroptosis in COPD

doi: 10.1016/j.redox.2024.103388

Figure Lengend Snippet: Fer-1 alleviated lipid peroxidation induced by CSE in Beas-2b cells. (A) CCK-8 assays indicated that Fer-1 reversed the CSE-induced death of Beas-2b cells. (B) LDH assays showed that Fer-1 suppressed LDH release caused by CSE. (C, D) The effects of Fer-1 on GSH/GSSG ratio and MDA production were determined by the corresponding kits. (E, F) The ability of Fer-1 to inhibit lipid peroxidation and ROS production was measured by Liperfluo and DCFH-DA. (G) Redox lipidomics was performed by LC-MS to differentiate different lipid peroxidation species in CSE- or Fer-1-treated cells. (H) The relative quantitative values of differential metabolites were normalized transformed and clustered. (I) The expression levels of xCT and GPx4 were measured by western blotting. (J) The mRNA levels of xCT and GPx4 were analyzed by qPCR. (K) Western blot showed Fer-1 failed to inhibit the degradation of xCT and GPx4. ∗p＜0.05,∗∗p＜0.01 compared with the control group; NS p＞0.05, #p＜0.05,##p＜0.01 compared with the CSE-treated group.

Article Snippet: Primary antibodies against caspase-4/11, FSP1, GCH1 and GPx4 were obtained from Zenbio (Chengdu, China).

Techniques: CCK-8 Assay, Liquid Chromatography with Mass Spectroscopy, Transformation Assay, Expressing, Western Blot, Control

Journal: Redox Biology

Article Title: Lipid peroxidation triggered by the degradation of xCT contributes to gasdermin D-mediated pyroptosis in COPD

doi: 10.1016/j.redox.2024.103388

Figure Lengend Snippet: Fer-1 protected against CS-induced inflammation and lipid peroxidation in the lungs. (A) Exudative inflammatory cells in BALF were examined by Giemsa staining and analyzed by optical microscopy (200 × ). (B) Pathologic changes were assessed by H&E staining (200 × ). (C) Mucus secretion by the airway epithelium was examined by PAS staining (200 × ). (D) The level of ROS in inflammatory cells was analyzed with a DCFH-DA probe and observed by fluorescence microscopy (200 × ). (E, F) The GSH and MDA contents were measured with detection kits. (G) The production of 4-HNE was quantified by immunohistochemical staining (200 × ). (H) The expression levels of xCT and GPx4 were assessed by western blotting. ∗∗p＜0.01 compared with the control group; #p＜0.05,##p＜0.01 compared with the CS-exposed group.

Article Snippet: Primary antibodies against caspase-4/11, FSP1, GCH1 and GPx4 were obtained from Zenbio (Chengdu, China).

Techniques: Staining, Microscopy, Fluorescence, Immunohistochemical staining, Expressing, Western Blot, Control

Journal: Redox Biology

Article Title: Lipid peroxidation triggered by the degradation of xCT contributes to gasdermin D-mediated pyroptosis in COPD

doi: 10.1016/j.redox.2024.103388

Figure Lengend Snippet: xCT downregulation contributed to lipid peroxidation and pyroptosis in Beas-2b cells. (A) The knockdown effects of three xCT-siRNAs were assessed by western blotting. (B) Cell viability was determined by a CCK-8 kit. (C, D) The levels of IL-1β and IL-18 were measured by ELISAs. (E, F) The contents of GSH and MDA were measured with corresponding kits. (G, H) Western blot analysis showed that xCT knockdown decreased the expression of components of the xCT/GPx4 axis and increased the expression of pyroptosis-related proteins. (I) Immunofluorescence staining was used to assess the expression of N-GSDMD, and the results were analyzed by immunofluorescence microscopy (200 × ). ∗p＜0.05,∗∗p＜0.01 compared with the respective control group; #p＜0.05,##p＜0.01 compared with CSE-exposed NC group.

Article Snippet: Primary antibodies against caspase-4/11, FSP1, GCH1 and GPx4 were obtained from Zenbio (Chengdu, China).

Techniques: Knockdown, Western Blot, CCK-8 Assay, Expressing, Immunofluorescence, Staining, Microscopy, Control

Journal: Redox Biology

Article Title: Lipid peroxidation triggered by the degradation of xCT contributes to gasdermin D-mediated pyroptosis in COPD

doi: 10.1016/j.redox.2024.103388

Figure Lengend Snippet: Cigarette smoke induced lipid peroxidation and pyroptosis through the ubiquitination and degradation of xCT. (A) Lipid peroxidation due to the suppression of the xCT/GPX4 axis facilitated pyroptosis in cigarette smoke-induced COPD. (B) The process of xCT protein ubiquitination. In an ATP-dependent manner, Ub is attached to the ubiquitin-activating enzyme E1 and then delivered to the ubiquitin-binding enzyme E2. Next, the E3 ligase acts as a bridge, identifying E2-Ub and the substrate xCT to facilitate the binding of Ub to xCT. Finally, the ubiquitin chain is identified by the 26S proteasome, leading to the degradation of xCT.

Article Snippet: Primary antibodies against caspase-4/11, FSP1, GCH1 and GPx4 were obtained from Zenbio (Chengdu, China).

Techniques: Binding Assay