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:

Journal: iScience

Article Title: Niche-expressed Galectin-1 is involved in pre-B acute lymphoblastic leukemia relapse through pre-B cell receptor activation

doi: 10.1016/j.isci.2023.106385

Figure Lengend Snippet: GAL1+ mesenchymal stromal cells favor the development of murine Pre-BCR+ B-ALL

Article Snippet: The expression of GAL1 was controlled with an anti mouse GAL1 antibody (R&D Systems) conjugated with the Alexa-Fluor 488 antibody labeling kit (Thermo Fisher).Patient samples and human cell lines.

Techniques:

Journal: iScience

Article Title: Niche-expressed Galectin-1 is involved in pre-B acute lymphoblastic leukemia relapse through pre-B cell receptor activation

doi: 10.1016/j.isci.2023.106385

Figure Lengend Snippet: Pre-BCR signaling and human pre-B ALL growth are impaired in the absence of GAL1

Article Snippet: The expression of GAL1 was controlled with an anti mouse GAL1 antibody (R&D Systems) conjugated with the Alexa-Fluor 488 antibody labeling kit (Thermo Fisher).Patient samples and human cell lines.

Techniques: