Journal: Cell Communication and Signaling : CCS

Article Title: PPARγ agonism ameliorates acute kidney injury by inhibiting neutrophil extracellular trap formation-mediated renal tubular epithelial cell PANoptosis

doi: 10.1186/s12964-026-02686-6

Figure Lengend Snippet: CP-induced NETosis occurs through a ROS-dependent pathway. Inhibition of neutrophil ROS reduces CP-induced NETosis. A Detection of Cit-H3 expression and assessment of neutrophil death levels using live/dead cell staining and immunofluorescence microscopy. Scale bar, 100 μm. B Western blot analysis of P62 and LC3B expression in neutrophils. C Kidney tissue sections were stained for 8-OHdG, and representative images with corresponding quantitative histograms are shown. In addition, frozen kidney tissue sections were stained with a ROS detection solution to visualize ROS levels. Scale bar, 50 μm. D Quantification of kidney ROS levels using a fluorescence microplate reader. E Representative fluorescence images showing ROS production (detected by DHE staining) in neutrophils. Scale bar, 200 μm. F Detection of ROS levels in bone marrow-derived neutrophils using a fluorescence microplate reader after stimulation with CP or PMA. G Fluorescence microscopy imaging of Cit-H3 expression in neutrophils treated with NAC or chloroquine followed by CP exposure. Scale bar: 50 μm. NAC: N-acetyl-L-cysteine (antioxidant); CQ: chloroquine (autophagy inhibitor). Experiments were performed in triplicate, and the results are presented as mean ± SEM. Statistical significance was analyzed using independent samples t-tests ( C , D ) or one-way ANOVA, followed by Tukey’s post hoc test ( B , F ). Significance levels: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns: not significant

Article Snippet: The antibody against 8-OHdG was purchased from Bioss (bs1278R, China).

Techniques: Inhibition, Expressing, Staining, Immunofluorescence, Microscopy, Western Blot, Fluorescence, Derivative Assay, Imaging