fluorescein  (Thermo Fisher)

Journal: Molecules

Article Title: Anti-Inflammatory and Immunomodulatory Effects of Aqueous Extracts from Green Leaves and Rhizomes of Posidonia oceanica (L.) Delile on LPS-Stimulated RAW 264.7 Macrophages

doi: 10.3390/molecules30244685

Figure Lengend Snippet: ( A ) Representative flow cytometric profiles of FITC-dextran uptake by RAW 264.7 cells cultured under control conditions, treated with 0.1 μg/mL LPS or co-treated with LPS and 10 μg/mL GLE or 0.1 μg/mL RE. Fully processed preparations without FITC-dextran were analyzed in parallel for the control of background derived from autofluorescence (negative control). ( B ) Bar graph showing the relative mean fluorescence intensity (MFI) of control (C), LPS, GLE + LPS, or RE + LPS-treated RAW 264.7 cells. All the results were analyzed for the MFI of each condition, and the MFI of either treated cell population was divided by the MFI of the controls to normalize the data. Error bars indicate the s.e.m. of three independent measurements. One-way ANOVA followed by Holm–Sidak comparison procedure was used. * p < 0.05 and ** p < 0.001 vs. LPS. Normality test vs. control passed.

Article Snippet: To quantify the effect of the co-treatments on the bulk-phase endocytic ability of the macrophages, RAW 264.7 cells were grown in control conditions or treated with LPS with or without addition of either 10 μg/mL GLE or 0.1 μg/mL RE for 24 h. Subsequently, the cells were incubated at 37 °C for 1 h with 100 μL of fluorescein isothiocyanate (FITC)-dextran (final concentration: 1 mg/mL; MW 40000; MedChem Express) and then subjected to flow cytometric analysis to evaluate their uptake, as described by Rod-In et al. [ ].

Techniques: Cell Culture, Control, Derivative Assay, Negative Control, Fluorescence, Comparison