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: Single‐cell transcriptomic analysis reveals cellular heterogeneity in the NC and TLE group. (A) UMAP visualization of the single‐cell RNA‐seq data from normal and TLE hippocampus samples, displaying diverse cell populations. The horizontal and vertical axes represent components after dimensionality reduction. Each point in the figure represents a cell, with cells in close proximity considered to be of the same type. Different cell groups are distinguished by different colors. (B) UMAP plots highlighting the distribution of 12 identified cell types in NC (four samples, left) and TLE groups (four samples, right). The horizontal and vertical axes represent components after dimensionality reduction. Each point in the figure represents a cell, with cells in close proximity considered to be of the same type. Different cell groups are distinguished by different colors. (C) Pie charts showing the proportion of different cell types in the NC (left) and TLE (right) groups, illustrating the changes in cellular composition. (D) Bar graph depicting the count or numbers of each cell type in NC and TLE groups. (E) Visualization of the expression of eight key genes ( Cdc25b , Dnmt1 , Fgd3 , Gzma , Mtx1 , Raf1 , Sh3bp5l , and Ssh2 ) across various cell types in the hippocampal tissue via a UMAP plot of gene expression. It is observed that, except for Fgd3 and Ssh2 which are specifically highly expressed in microglia, and Gzma in a small population of T cells, other genes do not exhibit cell type‐specific expression and are generally distributed across cell types. (F) Bubble chart of pathways significantly enriched in DEGs from single cell RNA sequence analysis of microglia comparing TLE and NC groups. Top 20 key pathways are listed on the y ‐axis, with the rich factor ( x ‐axis) indicating enrichment strength. Bubble size symbolize the count of genes involved, while color intensity reflects the significance of enrichment, with darker red indicating higher statistical significance. (G) KEGG pathway enrichment analysis while comparing microglia in TLE versus NC, for the target gene set ( Fgd3 and Ssh2 ). This KEGG pathway enrichment analysis delineates the regulation of the actin cytoskeleton, with a specific emphasis on the roles of GPCRs ( Bdkrb2 ), FGD1/3 ( Fgd3 ), IQGAP ( Iqgap1 ), Rac ( Rac2 ), PAK ( Pak2 ), SSH ( Ssh2 ), and the Arp2/3 ( Arpc1b ) complex. Arrow (→) indicates a promoting or activating effect. T‐shaped line (⊣) indicates an inhibitory or blocking effect. All entities with colored backgrounds in the diagram are part of the KEGG annotation results for genes/transcripts under TLE versus NC comparison. Yellow background indicates known genes/transcripts, while green background represents new genes/transcripts (none). Red borders denote upregulated genes, and blue borders indicate downregulated genes (none). Genes encircled in pink and blue ellipses are those for which protein expression validation was conducted subsequently (Figure E). Pink signifies that the corresponding gene's protein level in the TLE hippocampus is significantly upregulated, consistent with predictions, whereas blue indicates no significant upregulation was found. (H) Capillary‐based immunoblots showing the expression of key proteins upstream and downstream of Fgd3 and Ssh in the regulation of the actin cytoskeleton pathway. (I) Quantification of Western blot analysis of the protein bands, using relative chemiluminescence signal values compared across NC and TLE hippocampus samples. (* p < 0.05, ** p < 0.01, *** p < 0.001, ns, not significant). (TLE, temporal lobe epilepsy; NC, normal control.)

Article Snippet: For protein capillary electrophoresis, the primary antibodies used were IQGAP1 (#ab133490, 1:50, Abcam), BKRB2 (#YN2508; 1:50, Immunoway), PAK2 (#2608S, 1:50, CST), ARPC1B (#28368‐1‐AP, 1:50, Proteintech), and RAC2 (#10735‐1‐AP; 1:50, Proteintech).

Techniques: RNA Sequencing, Expressing, Gene Expression, Sequencing, Blocking Assay, Comparison, Biomarker Discovery, Western Blot, Control

Journal: PLoS ONE

Article Title: Exploring Regulatory Mechanisms of Atrial Myocyte Hypertrophy of Mitral Regurgitation through Gene Expression Profiling Analysis: Role of NFAT in Cardiac Hypertrophy

doi: 10.1371/journal.pone.0166791

Figure Lengend Snippet: Selected Signature Downregulated Gene Expressions in the Left Atria of Mitral Regurgitation vs. Normal Control

Article Snippet: ARPC3 , 10094 , actin related protein 2/3 complex, subunit 3 , BP: movement of cell or subcellular component, Arp2/3 complex-mediated actin nucleation, Fc-gamma receptor signaling pathway involved in phagocytosis, ephrin receptor signaling pathway, actin filament organization; CC: cytosol, Arp2/3 protein complex, actin cytoskeleton, extracellular vesicular exosome, cell projection, lamellipodium; MF: structural constituent of cytoskeleton, protein binding, actin filament binding , Regulation of actin cytoskeleton, Fc gamma R-mediated phagocytosis , -2.124.

Techniques: Binding Assay, Variant Assay, Activity Assay, Expressing, Migration, Protein Binding, RNA Binding Assay, Transduction, De-Phosphorylation Assay, Modification, Cell Differentiation, Activation Assay, Coagulation, Sequencing, Chemotaxis Assay, Sublimation, Histone Deacetylase Assay, Concentration Assay, Transferring, Translocation Assay