Journal: Frontiers in Cellular Neuroscience

Article Title: Differential Cellular Expression of Galectin-1 and Galectin-3 After Intracerebral Hemorrhage

doi: 10.3389/fncel.2019.00157

Figure Lengend Snippet: Increased expression of Galectin-1 after ICH. (A) Confocal images depicting the temporal expression pattern of Galectin-1 after ICH (scale bar = 20 μm; n = 3–5 per group). (B) The average number of Galectin-1-positive cells per 0.1 mm 2 in the ipsilateral striatum. (C) Increased expression of Galectin-1 was confirmed by western blotting of brain tissue from the ipsilateral striatum. (D) Densitometry analysis ( n = 3–5 per group) of the western blotting data. ∗ p < 0.05, ∗∗∗ p < 0.001 vs. sham.

Article Snippet: Cells were then incubated with mouse recombinant Galectin-1 (6.25 or 12.5 μg/ml; R&D Systems, Minneapolis, MN, United States) for 1 h and it was followed by an 18-h treatment with LPS (100 ng/ml) or hemin (30 μg/ml).

Techniques: Expressing, Western Blot

Journal: Frontiers in Cellular Neuroscience

Article Title: Differential Cellular Expression of Galectin-1 and Galectin-3 After Intracerebral Hemorrhage

doi: 10.3389/fncel.2019.00157

Figure Lengend Snippet: Galectin-1 expression is mostly observed in glial cells after ICH. Brain sections were double immunostained for (A) Galectin-1 and GFAP, (B) Galectin-1 and Iba1, (the lowest panel depicts the high magnification images) and (C) Galectin-1 and NeuN (Scale bar = 20 μm; n = 3 per group). Galectin-1 expression was observed mostly in GFAP-positive cells and in a subset of Iba1-positive cells. NeuN positive cells didn’t express Galectin-1 ( n = 3 per group).

Article Snippet: Cells were then incubated with mouse recombinant Galectin-1 (6.25 or 12.5 μg/ml; R&D Systems, Minneapolis, MN, United States) for 1 h and it was followed by an 18-h treatment with LPS (100 ng/ml) or hemin (30 μg/ml).

Techniques: Expressing

Journal: Frontiers in Cellular Neuroscience

Article Title: Differential Cellular Expression of Galectin-1 and Galectin-3 After Intracerebral Hemorrhage

doi: 10.3389/fncel.2019.00157

Figure Lengend Snippet: Recombinant Galectin-1 and inflammatory response. Prior to LPS (100 ng/ml) stimulation, Raw 264.7 cells were treated with recombinant Galectin-1, and the release of IL-6 was measured using ELISA, as detailed in methods. Recombinant Galectin-1 significantly reduced LPS-induced release of IL-6 from RAW 246.7 cells ( n = 4 per group). ∗∗∗ p < 0.001 vs. LPS.

Article Snippet: Cells were then incubated with mouse recombinant Galectin-1 (6.25 or 12.5 μg/ml; R&D Systems, Minneapolis, MN, United States) for 1 h and it was followed by an 18-h treatment with LPS (100 ng/ml) or hemin (30 μg/ml).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay

Journal: PLoS Pathogens

Article Title: A New Human 3D-Liver Model Unravels the Role of Galectins in Liver Infection by the Parasite Entamoeba histolytica

doi: 10.1371/journal.ppat.1004381

Figure Lengend Snippet: (A–D) ELISA quantification in compartments S1, S2 and S3 of the 3D-liver model (Model) or the setup without LSEC (no LSEC), in the absence or the presence of E. histolytica (− or + Eh). Samples used for ELISA assays were prepared as for . (A) Amounts of IL-6 at 3 h, (B) IL-1β, (C) galectin-1 and (D) galectin-3 at 6 h of incubation with fresh serum-free medium containing or not virulent E. histolytica . Graphs with standard deviation and statistical evaluation by One way ANOVA, p<0.01*, <0.001**, <0.0001*** for 3–5 independent experiments. None of the cytokines was detected in the COL-I matrix control without hepatic cells. n/d = not detected. (E, F) Summary of the results from and , for the setup without LSEC (E) and the 3D-liver model (F), showing the complexity of the pro-inflammatory response induced by E. histolytica in the 3D-liver model.

Article Snippet: For quantitative ELISA-like assays, trophozoites were harvested, allowed to adhere to wells of a 96 well plate (6×10 4 /well), incubated for 25 min with recombinant galectin-1, galectin-3 or BSA, gently washed and then incubated with biotinylated anti-galectin-1 or -3 antibodies (R&D systems).

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Standard Deviation

Journal: PLoS Pathogens

Article Title: A New Human 3D-Liver Model Unravels the Role of Galectins in Liver Infection by the Parasite Entamoeba histolytica

doi: 10.1371/journal.ppat.1004381

Figure Lengend Snippet: Bacterially expressed purified human galectin-1 and -3, and BSA as a control, were used for the incubations with the amoebae. Galectin-1 (A) or -3 (B) binding to the trophozoite surface, visualized by immunofluorescence with human galectin-specific antibodies. Quantification of galectin-1 (C) or -3 (D) binding to immobilized trophozoites by ELISA-like assays. (E) Trophozoite adhesion to immobilized galectin-1 or -3 and competition with lactose (lac).

Article Snippet: For quantitative ELISA-like assays, trophozoites were harvested, allowed to adhere to wells of a 96 well plate (6×10 4 /well), incubated for 25 min with recombinant galectin-1, galectin-3 or BSA, gently washed and then incubated with biotinylated anti-galectin-1 or -3 antibodies (R&D systems).

Techniques: Purification, Binding Assay, Immunofluorescence, Enzyme-linked Immunosorbent Assay

Journal: PLoS Pathogens

Article Title: A New Human 3D-Liver Model Unravels the Role of Galectins in Liver Infection by the Parasite Entamoeba histolytica

doi: 10.1371/journal.ppat.1004381

Figure Lengend Snippet: Immunofluorescence localization of cell surface-associated (A) galectin-3 on Huh-7 and (C) galectin-1 on LSEC. Amoeba adhesion to (B) Huh-7 or (D) LSEC 2D cultures Control (CTL), corresponding to incubations without proteins or sugars added, was set to 100%. (E) LSEC galectin-1 surface label 72 h after transfection with galectin-1 specific siRNA. (F) Amoeba binding to LSEC transfected with galectin-1 specific or unrelated control siRNA. Incubations and quantification of trophozoite binding as for B and D. Graphs with standard deviation and statistical evaluation by One way ANOVA, p<0.0001*** for 3 independent experiments.

Article Snippet: For quantitative ELISA-like assays, trophozoites were harvested, allowed to adhere to wells of a 96 well plate (6×10 4 /well), incubated for 25 min with recombinant galectin-1, galectin-3 or BSA, gently washed and then incubated with biotinylated anti-galectin-1 or -3 antibodies (R&D systems).

Techniques: Immunofluorescence, Transfection, Binding Assay, Standard Deviation

Journal: PLoS Pathogens

Article Title: A New Human 3D-Liver Model Unravels the Role of Galectins in Liver Infection by the Parasite Entamoeba histolytica

doi: 10.1371/journal.ppat.1004381

Figure Lengend Snippet: Cytokine quantification by ELISA assays in fractions S1 (A–C) or S3 (D) after 6 h of incubation of 3D-liver models with bacterially expressed purified human galectin-1 or -3 (1 µg/ml) or E. histolytica . Control (CTL) corresponds to no protein added. Incubations with amoebae were performed in the presence of lactose or in its absence (CTL). Graphs with standard deviation, statistical evaluation by One way ANOVA, p<0.05*, p<0.001**, p<0.0001*** for (A–C) and values significantly different from 0 for galectin-3 p = 0.0068, CTL p = 0,0049 (D), for 3 independent experiments. n/d = not detected.

Article Snippet: For quantitative ELISA-like assays, trophozoites were harvested, allowed to adhere to wells of a 96 well plate (6×10 4 /well), incubated for 25 min with recombinant galectin-1, galectin-3 or BSA, gently washed and then incubated with biotinylated anti-galectin-1 or -3 antibodies (R&D systems).

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Purification, Standard Deviation

Journal: PLoS Pathogens

Article Title: A New Human 3D-Liver Model Unravels the Role of Galectins in Liver Infection by the Parasite Entamoeba histolytica

doi: 10.1371/journal.ppat.1004381

Figure Lengend Snippet: Schematic presentation of the E. histolytica invasion mechanisms at the 3D-liver model: the early LSEC adhesion interactions and crossing as well as the induction of the pro-inflammatory cytokine release into the different compartments of the 3D-liver model. The summary highlights the involvement of amoebic Gal/GalNAc lectin in the interactions with the human cells, and the dual role of human galectin-1 and 3. First, surface-associated galectins promotes amoeba adhesion during the early stages, and second, released galectins participate in the induction of the inflammatory response later in the invasion process.

Article Snippet: For quantitative ELISA-like assays, trophozoites were harvested, allowed to adhere to wells of a 96 well plate (6×10 4 /well), incubated for 25 min with recombinant galectin-1, galectin-3 or BSA, gently washed and then incubated with biotinylated anti-galectin-1 or -3 antibodies (R&D systems).

Techniques: