Journal: Science Advances

Article Title: Cytoskeletal remodeling promotes tunneling nanotube formation and drives cardiac resident cell mitochondrial transfer in sepsis

doi: 10.1126/sciadv.adz3266

Figure Lengend Snippet: ( A ) Western blot analysis and Co-IP assay showing the Drp1 and Filamin/Kinesin binding ability of the myocardial tissue after CLP; n = 8 per group. ( B ) Functional pattern of the full-length domain of Drp1 protein. ( C and D ) Schematic diagram of the docking of Filamin and Kinesin with Drp1. ( E ) Immunofluorescence images showing the expression and distribution of Drp1 (green) and Filamin/Kinesin (red) in cardiomyocytes cocultured with endothelial cells from control and LPS-treated groups; n = 3 per group. Scale bars, 25 μm. ( F ) Immunofluorescence images showing the expression of Filamin/Kinesin (red) in cardiomyocytes cocultured with fibroblasts or macrophages; n = 3 per group. Scale bars, 25 μm. ( G and H ) Time-lapse recordings of mitochondrial transfer via TNT between WT and Drp1 eCKO primary cardiomyocytes and primary endothelial cells, fibroblasts, and macrophages by HIS-SIM. Mitochondria were labeled with PKmito (yellow); TNTs were labeled with tubulin (green) and actin (cyan); n = 3 per group. Scale bars, 10 μm. IgG, immunoglobulin G; GTPase, guanosine triphosphatase; aa, amino acids.

Article Snippet: Equal lysate amounts were incubated overnight with Drp1 antibody (1:50; Novus Biologicals, NB110-55288) or immunoglobulin G (1:50; Cell Signaling, #2729) at 4°C, followed by incubation with protein A/G magnetic beads (MedChemExpress, HY-K0202) for 6 hours.

Techniques: Western Blot, Co-Immunoprecipitation Assay, Binding Assay, Functional Assay, Immunofluorescence, Expressing, Control, Labeling

Journal: World Journal of Stem Cells

Article Title: GATA2 deficiency exacerbates chronic liver injury via disrupting hepatocyte death-regeneration balance: Clinical, histopathological, and molecular evidence

doi: 10.4252/wjsc.v18.i2.112940

Figure Lengend Snippet: Identification of GATA2 binding sites in gene promoters through chromatin immunoprecipitation sequencing and prediction of its regulatory functions. A: Scatter plot showing the correlation between biological replicates; B: Histogram depicting the distribution of peak lengths; C: Distribution of peaks across chromosomes (X-axis: Chromosome length; left Y-axis: Peak intensity; right Y-axis: Chromosome numbers); D: Read distribution surrounding transcription start sites (TSS) of peak-associated genes (top: Line chart of average sequencing depth around TSS; bottom: Heatmap of sequencing depth around TSS for peak-associated genes, ranked by average depth); E: Gene Ontology functional enrichment analysis of peak-associated genes; F: Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis of peak-associated genes; G: Fold enrichment of selected genes associated with partial regeneration, molecular chaperones, antioxidant response, and fatty acid β-oxidation as determined by chromatin immunoprecipitation sequencing; H: Validation of GATA2 binding to the promoters of heat shock protein family D member 1, glutathione peroxidase 4, and peroxisome proliferator-activated receptor alpha by chromatin immunoprecipitation real-time quantitative polymerase chain reaction. Data are presented as fold enrichment relative to immunoglobulin G; I: Western blot analysis of GATA2, heat shock protein 60, glutathione peroxidase 4, and peroxisome proliferator-activated receptor alpha protein expression following transfection with the Gata2 construct. b P < 0.01 vs immunoglobulin G or control group. ChIP-seq: Chromatin immunoprecipitation sequencing; IgG: Immunoglobulin G; PCNA: Proliferating cell nuclear antigen; HSPD1: Heat shock protein family D member 1; GPX4: Glutathione peroxidase 4; UCP2: Uncoupling protein 2; PPARA: Peroxisome proliferator-activated receptor alpha.

Article Snippet: An additional 10% was incubated with rabbit immunoglobulin G (IgG) (Cell Signaling Technology, Cat# 2729, Danvers, MA, United States) as a negative control and labeled as “IgG”.

Techniques: Binding Assay, ChIP-sequencing, Sequencing, Functional Assay, Biomarker Discovery, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Western Blot, Expressing, Transfection, Construct, Control