Journal: Cell Death Discovery

Article Title: Pregnane X receptor protects against age-related bone loss in males via PI3K/Akt-mediated inhibition of apoptosis

doi: 10.1038/s41420-025-02797-y

Figure Lengend Snippet: A Schematic diagram of experimental design. B ELISA for inflammation factors Tnfα, Il1β, and Il6 in the conditional medium collected from Nr1i2 knockdown primary BMSCs or siNC as control. C ELISA for ROS clearance-related enzyme T-SOD, GSH-PX and ROS damage biomarker MDA in Nr1i2 knockdown primary BMSCs or siNC as control. D Flow cytometry analysis of intracellular ROS level by using the fluorescent dye DCFDA. Western blot for PI3K/Akt pathway- ( E ) and apoptosis- ( F ) related proteins in primary BMSCs of different groups. Data were means ± s.e.m. n = 3 independent repeats for in vitro experiments. ** p < 0.01, *** p < 0.001 by t-test.

Article Snippet: The Il1β, Il6, and Tnfα level in the conditional medium, intracellular level of T-SOD, GSH-PX, MDA, and ROS were determined by using the Mouse IL-1β ELISA Kit (HJ177, Epizyme Biotech, Shanghai, China), Mouse IL-6 ELISA Kit (HJ182, Epizyme Biotech, Shanghai, China), and Mouse TNF-α ELISA Kit (HJ207, Epizyme Biotech, Shanghai, China), Total superoxide dismutase (SOD) assay kit (A001-3-2, Jiancheng, Nanjing, China), Glutathione Peroxidase (GSH-PX) Assay Kit (A005-1-2, Jiancheng, Nanjing, China), Malondialdehyde (MDA) assay kit (A003-1-2, Jiancheng, Nanjing, China), and DCFDA/H2DCFDA-Cellular ROS Assay Kit (ab113851, abcam, Cambridge, UK) according to the manufacturer’s instructions, respectively.

Techniques: Enzyme-linked Immunosorbent Assay, Knockdown, Control, Biomarker Discovery, Flow Cytometry, Western Blot, In Vitro