Journal: International Journal of Molecular Sciences

Article Title: eDNA, Amyloid Fibers and Membrane Vesicles Identified in Pseudomonas fluorescens SBW25 Biofilms

doi: 10.3390/ijms232315096

Figure Lengend Snippet: P. fluorescens biofilms include amyloid fibers. Confocal laser scanning microscopy (CLSM) was used to visualize amyloid fibers in three-day-old Viscous Mass and Wrinkly Spreader and biofilms. Shown here are single-channel and combined CLSM images of Viscous Mass ( A ) and Wrinkly Spreader ( B ) three-day-old biofilms. AmyGreen (green channel) was used to visualize amyloid fibers, Calcofluor white (blue channel) to visualize cellulose, and ethidium bromide (red channel) to visualize total DNA as a proxy for live cells, respectively, and all three channels are combined in the bottom-right images. Note the low signal region running down the Wrinkly Spreader image. The dimensions of the images are 115 × 115 × 20/35 µm.

Article Snippet: Intracellular DNA from intact and metabolically active (live) cells was stained with 5 μM SYTO 9 solution (ThermoFisher Scientific), and total DNA (eDNA and DNA in active and dead cells) stained with 2 μg/mL ethidium bromide (ThermoFisher Scientific).

Techniques: Confocal Laser Scanning Microscopy

Journal: Signal transduction and targeted therapy

Article Title: The cGAS-STING pathway-dependent sensing of mitochondrial DNA mediates ocular surface inflammation.

doi: 10.1038/s41392-023-01624-z

Figure Lengend Snippet: Fig. 4 mtDNA leakage into the cytosol mediated the cGAS-STING pathway activation under hyperosmotic stress. a Representative fluorescent micrographs of Ctrl and HS groups stained with dsDNA (green) and mitochondria (red). Image on the lower portion represents an enlarged section of the square frame in the upper portion. Scale bars: 10 μm. b Cytoplasmic mtDNA copy number was determined by real-time PCR (n = 5). c, d siRNA-mediated STING knockdown blocked mtDNA-induced p-TBK1 and p-IRF3 (n = 3). e Cytoplasmic mtDNA copy number was determined by real-time PCR after treatment for 48 h with EtBr (0.2 mg/ml) (n = 6). f–h HS-induced activation of STING, p-TBK1 and p-IRF3 and upregulation of CXCL10 and IFN-β were reversed by EtBr-mediated mtDNA depletion (n = 3 or 6). i, j 8-OHDG did not ameliorate HS-induced increases in STING levels or p-TBK1 and p-IRF3. The data were shown as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001

Article Snippet: Signal Transduction and Targeted Therapy (2023) 8:371 Reagent and antibodies C-176, ethidium bromide (EtBr) and cyclosporin A (CsA) were obtained from Selleck (Houston, TX, USA).

Techniques: Activation Assay, Staining, Real-time Polymerase Chain Reaction, Knockdown