Journal: Molecular Neurobiology
Article Title: Exosome-Derived lncRNA NEAT1 Exacerbates Sepsis-Associated Encephalopathy by Promoting Ferroptosis Through Regulating miR-9-5p/ TFRC and GOT1 Axis
doi: 10.1007/s12035-022-02738-1
Figure Lengend Snippet: Exosome carried NEAT1 into the cerebral cortex and NEAT1 was significantly highly expressed in sepsis-induced ferroptosis A Brain vascular permeability was detected by EBD leakage in control and model rats ( n = 10). Extracted dye contents in the formamide extracts were quantified at 620 nm; B ROS level in cerebral cortex homogenate was detected by flow cytometry. C – E ELISA analysis on Fe ion, GSH, and GPX4 levels. F HE staining on cerebral cortex in control and model rats. G Western blot analysis on serous exosome biomarkers of TSG01, CD9, and CD63. H , I The expression of NEAT1 was detected by qRT-PCR in the serous exosome (normalized to synthetic cel-miR-39-3p) and cerebral cortex (normalized to GAPDH), respectively. J The correlation analysis on the expression of NEAT1 in exosome and cerebral cortex. ** P < 0.01 vs Control; *** P < 0.001 vs Control; EBD, Evans blue dye.
Article Snippet: Subsequently, the samples was incubated with primary antibodies, including rabbit anti-mouse polyclonal antibodies to TSG01 (1:2000; ab125011), CD9 (1:2000; ab92726), CD63 (1:2000; ab193349), TFRC (1:2000; ab214039), GOT1 (1:1000; ab221939) and GAPDH (1:5,000; ab8245) for 1 h. The samples were then incubated in the secondary antibody, horseradish peroxidase (HRP)-labeled goat anti-rabbit IgG (1:20,000, BA1054, BOSTER Inc.) with oscillation at room temperature for 40 min.
Techniques: Permeability, Control, Flow Cytometry, Enzyme-linked Immunosorbent Assay, Staining, Western Blot, Expressing, Quantitative RT-PCR
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