Journal: bioRxiv

Article Title: Transketolase of Staphylococcus aureus is involved in the control of master regulators of stress response during infection

doi: 10.1101/538900

Figure Lengend Snippet: A, B. Intracellular proliferation of USA300 wild type (WT) and Δ tkt mutant (Δ tkt ) strains. Endothelial EA.Hy926 cells expressing mKate2 nuclear restricted red fluorescent protein were infected with GFP expressing WT and Δ tkt strains. One hour after infection, cells were washed several times with gentamicin containing medium to eliminate extracellular bacteria. Gentamicin concentration of 50µg/ml was maintained throughout the experiment. Images were acquired every 30 minutes using Incucyte ® S3 live cell imaging system. A. Representative images (see also video S1 and S2). Red: cell nuclei. Green: GFP expressing bacteria. For each line, a white circle was centred on a unique infected cell. Bar = 100µm. B. Quantitative analysis representing the number of green particles divided by the number of red nuclei during the first day after infection. Blue line: WT strain. Red line: Δ tkt strain. Black line: non infected cells. C. Persistence of intracellular WT and Δ tkt strains. Endothelial EA.Hy926 cells were infected with WT and Δ tkt strains as described above. Gentamicin was maintained throughout the experiment. Survival of S. aureus was assessed by plating and counting CFUs three, seven and ten days after infection. Results are shown as mean ± SD of triplicate measurements.

Article Snippet: Plates were incubated and monitored at 5% CO2 and 37°C for 24 hours in the fully automated microscope Incucyte ® S3 (Essen BioScience).

Techniques: Mutagenesis, Expressing, Infection, Bacteria, Concentration Assay, Live Cell Imaging

Journal: BMC Complementary Medicine and Therapies

Article Title: Anti-inflammatory effect of Salsola komarovii extract with dissociated glucocorticoid activity

doi: 10.1186/s12906-020-02979-4

Figure Lengend Snippet: Salsola komarovii ethanol extract (SEE) as a glucocorticoid receptor (GR) ligand induces nuclear GR translocation. Binding affinity of a SEE and b hydrocortisone (HC) to the GR. Data are shown as fluorescence polarization (mP). The results are presented as mean ± standard error of the mean (SEM) of three independent experiments. c Immunofluorescence analysis of GR translocation into nucleus by HC or SEE. HaCaT cells were incubated for 24 h with 1 μM HC (positive control) or 40 μg/mL SEE followed by staining of GR (red fluorescence) and nucleus with 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI, blue fluorescence); original magnification 400×; scale bar = 100 μm

Article Snippet: After 3 h of incubation 24 °C, fluorescence polarization (FP) was measured using the 2103 EnVision multi-label plate reader (PerkinElmer) at 531 nm excitation and 595 nm emission wavelengths.

Techniques: Translocation Assay, Binding Assay, Fluorescence, Immunofluorescence, Incubation, Positive Control, Staining

Journal: STAR Protocols

Article Title: Isolation of murine hepatic myeloid cells with high yield and purity using immunomagnetic beads for subset analysis

doi: 10.1016/j.xpro.2023.102592

Figure Lengend Snippet:

Article Snippet: Centrifuge the sample for 3 min at 16,000 × g at 20°C–24°C in benchtop centrifuge (Thermo Fisher Scientific, #75002421).

Techniques: Recombinant, Plasmid Preparation, Blocking Assay, Purification, Control, Staining, Hood, Software