Journal: Bone Research
Article Title: DNMT aberration-incurred GPX4 suppression prompts osteoblast ferroptosis and osteoporosis
doi: 10.1038/s41413-024-00365-1
Figure Lengend Snippet: Mouse osteoporotic femurs induced by ovariectomy display marked ferroptosis and GPX4 suppression. a Representative microphotographs of H&E-stained distal femur sections (upper panel, arrows indicated trabeculae) and micro-CT-scanned (μ-CT) and 3-D reconstructed distal femurs from sham or Ovx mice (lower panel). Female C57BL/6 J mice of 12 weeks old were subjected to sham or ovariectomy (Ovx) surgery for 6 weeks ( n = 6). b Quantitative analysis of μ-CT data in ( a ) for bone volume to tissue volume ratio (BV/TV), trabecular number (Tb.N), trabecular thickness (Tb.Th) and trabecular separation (Tb.Sp), presented as box-and-whisker plots. * P < 0.05, n = 6, Student’s t test. c Representative distal femur sections stained by Perls’ method (upper panel, arrows indicate iron deposition) and by TUNEL staining (lower panel, arrows indicate positively-stained cells). d Western blotting. Femur homogenates from Sham and Ovx mice, control (Ctrl) and OP patients were assayed for type 1 collagen (Col 1), NFATc1, GPX4, 4-HNE and FSP1. GAPDH served as control. Three randomly-selected samples from 6 in each group were shown. Quantification of ( d ) on the right side of the blots was presented as mean ± SEM. * P < 0.05, Student’s t test. e Volcano plot of gene expression profile from database CRA007214 (total 21 422 genes, n = 3 mice for Sham and Ovx group). The number and position of genes statistic-significantly increased (2 535, red), no difference (15 573, gray) or decreased (3 314, blue) including Gpx4 (Log 2 fold change, FC = –2.499 06, yellow point) were marked. f Representative distal femur sections of Sham and Ovx mice stained for GPX4 by immunohistochemical (IHC) staining. The images on the right side of each panel were enlarged frames of the left panel. Positively-stained cells in brown were indicated by arrows. g Representative TEM micrographs of the femurs of Sham and Ovx mouse. Normal and ferroptotic mitochondria were indicated by blue and red arrows, respectively
Article Snippet: Perl’s Prussian blue staining (G1029, Servicebio, china) and TUNEL (TdT-mediated dUTP Nick-End Labeling, A111-01, Vazyme, China) followed the instructions in the kits.
Techniques: Staining, Micro-CT, Whisker Assay, TUNEL Assay, Western Blot, Control, Gene Expression, Immunohistochemical staining, Immunohistochemistry
