Journal: Frontiers in Microbiology

Article Title: Lacticaseibacillus casei Strain T21 Attenuates Clostridioides difficile Infection in a Murine Model Through Reduction of Inflammation and Gut Dysbiosis With Decreased Toxin Lethality and Enhanced Mucin Production

doi: 10.3389/fmicb.2021.745299

Figure Lengend Snippet: The Lacticaseibacillus casei -conditioned medium (LCM) of L. casei T21 attenuated C. difficile -induced IL-8 production, host gene expression, and increased transepithelial electrical resistance of C. difficile -stimulated colonic epithelial cells. The results are shown as IL-8 production in Caco-2 and HT-29 cells, respectively (A,B) ; IL-8 gene expression (relative to GAPDH ) in Caco-2 and HT-29 cells, respectively (C,D) ; the expression of SLC11A1 , HuR , and MUC-2 in Caco-2 cells, respectively (E–G) ; the expression of SLC11A1 , HuR , and MUC-2 in HT-29 cells, respectively (H–J) ; the transepithelial electrical resistance (TEER) values of Caco-2 cells (K) ; and the pH of cell culture medium (McCoy’s 5A modified medium for HT-29 cells) (L) . The results were from three independent experiments each in triplicate and expressed as the mean ± SEM. *p < 0.05.

Article Snippet: Colonic epithelial cells were then incubated with viable cells of C. difficile ATCC BAA1870 at multiplicity of infection (MOI) 1:300 either alone or with 5% (vol/vol) LCM for 24 h in 5% CO 2 at 37°C.

Techniques: Gene Expression, Expressing, Cell Culture, Modification

Journal: Advanced Science

Article Title: A Frog Skin‐Derived Peptide Targeting SCD1 Exerts Radioprotective Effects Against Skin Injury by Inhibiting STING‐Mediated Inflammation

doi: 10.1002/advs.202306253

Figure Lengend Snippet: Macropinocytosis‐mediated uptake of radiation‐induced frog skin peptide 2 (RIFSP‐2) protects skin cells against radiation‐induced injury. A) Administration of FITC‐labeled RIFSPs to HaCaT cells and identification of the membrane impermeability of RIFSP‐2 through confocal microscopy. B) Time‐ and dose‐dependent uptake of FITC‐RIFSP‐2 by HaCaT cells detected through flow cytometry analysis. C) Influence of heparin, amiloride, and cytochalasin D on the uptake of FITC‐RIFSP‐2 by HaCaT cells through confocal microscopy detection. The effect of RIFSP‐2 on D) cell proliferation, E) cell cycle, and F,G) senescence, as detected by colony formation assay, propidium iodide (PI) staining and SA‐β‐Gal staining analysis, respectively. H) Colocalization analysis showing FITC‐RIFSP‐2 (green) and Mito‐Tracker (red) in irradiated HaCaT and WS1 cells. I–L) Influence of RIFSP‐2 on cellular ROS accumulation, mitochondrial ROS accumulation, energy production, and mitochondrial membrane potential in irradiated WS1 cells through DCFH‐DA staining, MitoSox probe staining, ADP probe staining and JC‐1staining, respectively. M) Effect of RIFSP‐2 on the homeostasis of actin filaments in irradiated skin cells, as detected by rhodamine phalloidin and G‐actin staining assays. N) The actin polymerization inhibitor, cytochalasin B (CytB, 20µ m , 2h), and the actin depolymerization agonist, 5a‐Pregnane‐3,20‐dione (5αP, 100n m , 24h) were administrated before RIFSP‐2 treatment. Investigation of cell death in irradiated skin cells was adopted with AV/PI staining kits. O) and P) Calcein AM release assay for understanding the influence of RIFSP‐2 on lytic cell death in irradiated HaCaT cells. * P < 0.05 and ** P < 0.01, compared with the control group.

Article Snippet: The WS1 cell line (human skin fibroblasts) was purchased from ATCC.

Techniques: Labeling, Membrane, Confocal Microscopy, Flow Cytometry, Colony Assay, Staining, Irradiation, Release Assay, Control