Journal: CNS Neuroscience & Therapeutics

Article Title: Integrated Mendelian Randomization and Single‐Cell Transcriptomics Analysis Identifies Critical Blood Biomarkers and Potential Mechanisms in Epilepsy

doi: 10.1111/cns.70172

Figure Lengend Snippet: Expression of key genes at mRNA and protein levels in the hippocampus of TLE animal model. (A) Construction of the TLE mouse model. It primarily illustrates the process of animal anesthesia and fixation, kainic acid injection localization, brain tissue separation on ice, and hippocampus extraction. (B) Expression levels of core genes' corresponding mRNA in the hippocampus of NC and TLE groups identified by qPCR. Data are shown as mean ± SE. (C) Western blot analysis of hippocampal lysates displaying the protein levels of Dnmt1, Cdc25b, Ssh2, Fgd3, Gzma, Raf1, and Mx1 with Actin serving as a loading control. Approximate molecular weights are indicated. (D) Quantification of Western blot analysis of the protein bands in Figure C, through relative gray values compared across NC and TLE hippocampus samples. (E) Quantitative analyses of immunofluorescence staining showing the fluorescence intensity between NC and TLE in Figure F. (F) Representative immunofluorescence images showing the hippocampal localization of Cdc25b, Raf1, Fgd3, Dnmt1, Mx1, and Ssh2 (red) with DAPI staining the nuclei (blue) in NC and TLE groups (Scale bar, 100 μm). (TLE, temporal lobe epilepsy; NC, normal control. * p < 0.05, ** p < 0.01, *** p < 0.001, ns, not significant.)

Article Snippet: Subsequently, slices were briefly rinsed in PBS for 5 min and stained with the DAPI nuclear marker (# D1306, 1:10000, Molecular Probes) for 5 min. After a final PBS rinse, slices were mounted on gel‐coated slides, cover‐slipped with Prolong gold anti‐fade (# P36930, Vector Laboratories), and stored at 4°C until further analysis.

Techniques: Expressing, Animal Model, Injection, Extraction, Western Blot, Control, Immunofluorescence, Staining, Fluorescence