Journal: Neurochemical Research
Article Title: Reactive Astrocytes Release GDNF to Promote Brain Recovery and Neuronal Survival Following Ischemic Stroke
doi: 10.1007/s11064-025-04370-6
Figure Lengend Snippet: Astrocyte-derived GDNF reduced neuronal death after OGD. (a , b) WB images and analysis of GDNF expression in primary cultured astrocytes (a) and GDNF content in the medium (b) at different conditions. N = 4 independent experiments. For OGD condition, primary cultured astrocytes were subjected to 6 h OGD and followed by 24 h reperfusion for western blot and ELISA analysis. (c) Schematic diagram showing experimental design. Primary astrocytes with or without transfection of GDNF plasmid were subjected to 6 h OGD. Astrocyte conditioned medium from astrocyte cultures not subjected or subjected to OGD, namely ACM and ACM(OGD) of different conditions were collected 24 h later and added to primary neuronal cultures after subjected to 1 h OGD followed by 24 h reoxygenation for neuronal viability and death assays. (d-f) Neuronal viability, percentages of PI + neurons and neurons with condensed nuclei after 1 h OGD and followed by 24 h reoxygenation at different conditions. The percentage of PI + neurons was calculated based on the total number of neurons based on DAPI staining. (g) Fluorescent images of neurons stained with PI and Dapi at different conditions. Data in (d) were averaged values from 6 replicates of representative experiment; data in (e , f) were averaged from cell counting of 9 images in each condition. * p < 0.05, ** p < 0.01, *** p < 0.001; (one-way ANOVA test)
Article Snippet: The primary antibodies include a mouse anti-GFAP monoclonal antibody (1:300; Cat. No. MAB360, Sigma), a rabbit anti-GDNF polyclonal antibody (1:300, Cat. No. SC-328, Santa Cruz Biotechnology), a rabbit anti-Iba 1 polyclonal antibody (1:300, Cat. No. 019-19741, Fujifilm).
Techniques: Derivative Assay, Expressing, Cell Culture, Western Blot, Enzyme-linked Immunosorbent Assay, Transfection, Plasmid Preparation, Staining, Cell Counting