dna probe dapi  (Danaher Inc)

Journal: Stem Cell Research & Therapy

Article Title: Human MuStem cells repress T-cell proliferation and cytotoxicity through both paracrine and contact-dependent pathways

doi: 10.1186/s13287-021-02681-3

Figure Lengend Snippet: Expression and secretion of immunoregulatory mediators in cultured human MuStem cells and bone marrow-derived mesenchymal stem cells. a Flow cytometry comparison of heme oxygenase-1 (HO-1) expression in hMuStem cells and BM-MSCs. b Fluorescent immunolabeling of inducible nitric oxide synthase (iNOS) in hMuStem cells and BM-MSCs. Lipopolysaccharide (LPS)-human monocyte-derived activated macrophages for 24 h and RAW 264.7 cell line were used as positive (C+) and negative (C−) controls, respectively. Nuclei were counterstained with DAPI (blue). Scale bars, 100 µm. c Representative RT-PCR profile of indoleamine 2,3-dioxygenase-1 (IDO-1) gene obtained for hMuStem cells and BM-MSCs. For each sample, the level of IDO-1 expression was quantified using the average mRNA level of IDO-1 obtained in unstimulated BM-MSCs as a reference. LPS-human monocyte-derived activated macrophages for 24 h and water were used as positive (C+) and negative (C−) controls, respectively. d Interleukin, growth factor, enzyme and carbohydrate-binding protein secretion profile of hMuStem cells and BM-MSCs. ELISA assays were performed using culture supernatant collected 24 h after medium change. Results are expressed as individual values and normalized as concentration relative to 1 million cultured cells (ng or pg/10 6 cells). Each experiment was performed on at least 5 and 4 independent batches of hMuStem cells and BM-MSCs, respectively. Stimulation corresponds to a 24-h treatment with 50 ng/mL of TNF-α and IFN-γ. * p < 0.05, ** p < 0.01, ** p < 0.001, **** p < 0.0001; Wilcoxon matched-pairs signed rank test (unstimulated vs. stimulated) or Mann–Whitney U test (hMuStem cells vs. BM-MSCs). US, unstimulated; S, TNF-α/IFN-γ-stimulated

Article Snippet: After incubation (1 h, RT) in blocking buffer (5% goat serum in PBS), cells were incubated overnight with iNOS Ab (1:100, sc-651 clone, Santa Cruz Biotechnology, Santa Cruz, CA, USA) and counterstained (15 min, 37 °C) with DAPI fluorescent cell-permeable DNA probe (Life Technologies Ltd, Paisley, UK).

Techniques: Expressing, Cell Culture, Derivative Assay, Flow Cytometry, Comparison, Immunolabeling, Reverse Transcription Polymerase Chain Reaction, Binding Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Biosorption of Pb (II) and Zn (II) from aqueous solution by Oceanobacillus profundus isolated from an abandoned mine

doi: 10.1038/s41598-020-78187-4

Figure Lengend Snippet: Confocal images of O. profundus KBZ 3-2 cultured in 20 mg/L Pb for 24 h; (a) bright-field image, (b) bacterial cells stained by DAPI (blue), (c) EPS stained by Alexa Fluor 633-conjugated agglutinin (red), (d) Pb (II) stained by Leadmium Green AM Dye (green), (e) overlay image of (b–d ).

Article Snippet: Three different staining dyes were sequentially added to the cell suspension to stain the DNA (DAPI Nucleic Acid Stain, Molecular probes, Invitrogen), EPS (Wheat Germ Agglutinin, Alexa Fluor 633 Conjugate, Molecular Probes, Invitrogen), and Pb (II) (Leadmium Green AM Dye, Molecular Probes, Invitrogen).

Techniques: Cell Culture, Staining