Journal: Pediatric research

Article Title: Galectin-3 Expression and Effect of Supplementation in Neonatal Mice with Disseminated Candida albicans Infection

doi: 10.1038/s41390-019-0279-x

Figure Lengend Snippet: Tissue fungal burden and galectin-3 concentrations in adult mice with disseminated candidiasis. Mice (n=7) were infected via tail-vein injection with C. albicans and euthanized at 48 h after injection. Panel A: Tissue fungal burden. Panel B: Mean gal3 concentration in tissue homogenates compared to animals receiving saline (n=10). Error bars represent standard deviation. No differences in tissue gal3 concentration were detected by ANOVA with inter-group comparisons by the Student-Newman-Keuls Method.

Article Snippet: In a subsequent experiment, pups were delivered and randomized to receive 5 μg carrier-free recombinant mouse gal3 (R&D Systems, Minneapolis, MN) or saline in 20 μl i.p. injection prior to infection with C. albicans as described above.

Techniques: Infection, Injection, Concentration Assay, Saline, Standard Deviation

Journal: Pediatric research

Article Title: Galectin-3 Expression and Effect of Supplementation in Neonatal Mice with Disseminated Candida albicans Infection

doi: 10.1038/s41390-019-0279-x

Figure Lengend Snippet: Tissue fungal burden and galectin-3 concentrations in neonatal mice with disseminated candidiasis. Two-day-old mouse pups (n=14) were infected via intraperitoneal injection with C. albicans and euthanized at 24 h after injection. Panel A: Tissue fungal burden. Panel B: Mean gal3 concentration in tissue homogenates compared to pups receiving saline (n=14). Error bars represent standard deviation. Mean gal3 concentration was reduced in the spleen of infected pups relative to controls based on ANOVA with inter-group comparisons by the Student-Newman-Keuls Method (p=0.04).

Article Snippet: In a subsequent experiment, pups were delivered and randomized to receive 5 μg carrier-free recombinant mouse gal3 (R&D Systems, Minneapolis, MN) or saline in 20 μl i.p. injection prior to infection with C. albicans as described above.

Techniques: Infection, Injection, Concentration Assay, Saline, Standard Deviation

Journal: Pediatric research

Article Title: Galectin-3 Expression and Effect of Supplementation in Neonatal Mice with Disseminated Candida albicans Infection

doi: 10.1038/s41390-019-0279-x

Figure Lengend Snippet: Survival curve of neonatal mice with disseminated candidiasis after pretreatment with recombinant galectin-3. Two-day-old mouse pups were given intraperitoneal injections of either saline or recombinant gal3, 2 h prior to infection with C. albicans . Uninfected pups receiving gal3 only were included as a control. Pretreatment with gal3 reduced mortality in infected compared to saline treated pups based on log-rank test (p=0.02).

Article Snippet: In a subsequent experiment, pups were delivered and randomized to receive 5 μg carrier-free recombinant mouse gal3 (R&D Systems, Minneapolis, MN) or saline in 20 μl i.p. injection prior to infection with C. albicans as described above.

Techniques: Recombinant, Saline, Infection, Control

Journal: Pediatric research

Article Title: Galectin-3 Expression and Effect of Supplementation in Neonatal Mice with Disseminated Candida albicans Infection

doi: 10.1038/s41390-019-0279-x

Figure Lengend Snippet: Tissue fungal burden in neonatal mice with disseminated candidiasis pretreated with galectin-3. Two-day-old mouse pups were given intraperitoneal injections of either saline or recombinant gal3, 2 h prior to infection with C. albicans (n=10 pups per group). Tissues were collected at the time of death or at 72 h in surviving animals. Fungal burden is depicted with the bars representing median values. P values were derived based on analysis using a negative binomial model to account for the variability in these data which are not normally distributed.

Article Snippet: In a subsequent experiment, pups were delivered and randomized to receive 5 μg carrier-free recombinant mouse gal3 (R&D Systems, Minneapolis, MN) or saline in 20 μl i.p. injection prior to infection with C. albicans as described above.

Techniques: Saline, Recombinant, Infection, Derivative Assay

Journal: Pediatric research

Article Title: Galectin-3 Expression and Effect of Supplementation in Neonatal Mice with Disseminated Candida albicans Infection

doi: 10.1038/s41390-019-0279-x

Figure Lengend Snippet: Tissue fungal burden in neonatal mice with disseminated candidiasis at early time points following infection and pretreatment with galectin-3. Two-day-old mouse pups were given intraperitoneal injections of either saline or recombinant gal3, 2 h prior to infection with C. albicans (n=5 pups per group). Pups were euthanized and tissues were collected at 24 and 36 h following infection. Fungal burden is depicted with the bars representing median values. P values were derived based on analysis using a negative binomial model to account for the variability in these data which are not normally distributed. NS – not significant.

Article Snippet: In a subsequent experiment, pups were delivered and randomized to receive 5 μg carrier-free recombinant mouse gal3 (R&D Systems, Minneapolis, MN) or saline in 20 μl i.p. injection prior to infection with C. albicans as described above.

Techniques: Infection, Saline, Recombinant, Derivative Assay

Journal: BMC Women's Health

Article Title: The relationship between serum galectin-3 level and echocardiographic findings in patients with polycystic ovary syndrome

doi: 10.1186/s12905-026-04309-1

Figure Lengend Snippet: Scatter plot diagram between galectin-3 and global longitidunal strain

Article Snippet: Serum galectin-3 concentrations were calculated from a standard curve generated using the recombinant galectin-3 standards provided in the kit.

Techniques:

Journal: Nature Communications

Article Title: Secretory GFP reconstitution labeling of neighboring cells interrogates cell–cell interactions in metastatic niches

doi: 10.1038/s41467-023-43855-2

Figure Lengend Snippet: a t-Distributed stochastic neighbor embedding (t-SNE) plot of the GFP- hepatocytes from the healthy liver (control, n = 87) or the metastasized liver (distal, n = 59) and GFP+ hepatocytes from the metastasized liver (proximal, n = 99). Marker genes (two-sided version of Wilcoxon Rank-Sum test, adjusted p -value < 0.1, Log 2 FC > 0.25) among hepatocytes (proximal, distal, control) were used for clustering. Inserted numbers indicate cluster identification (left panel). The fraction rate of the hepatocytes in each cluster identified in the t-SNE plot (right panel). b Heatmap displaying expression level of clusters 2 and 3 marker genes of the top 20 ranked by Log 2 FC (two-sided version of Wilcoxon Rank-Sum test, adjusted p -value < 0.05) expressions in the GFP+ proximal hepatocytes. c The expression level of Lgals3 in the proximal, distal, and control. Data were statistically analyzed with analysis of variance and Tukey honestly significant difference test (95% family-wise confidence level). The adjusted p -values are shown in the graph. d Representative immunofluorescent staining images of galectin-3 in a metastasized liver section. Cancer cells (H2B-Azurite), hepatocytes (H2B-mCherry), and galectin-3 signals in the area of the hepatic tissue–metastasized colonies border (Liver–Meta border), distal liver tissue (Distal liver) and non-peripheral area of the metastatic colony (Non-peri meta) are shown. Similar results were observed in independent duplicate experiments. A scale bar indicates 20 µm. e Tumor spheroid proliferation assay of E0771 cells treated with galectin-3 in three-dimensional culture. Data were statistically analyzed with Holm-Sidak adjusted multiple t -test ( n = 102, 93, 90 and 72 for 0, 3, 5, 10 µg/mL galectin-3 respectively). The p -values are indicated in the graph. Data are presented as mean values ± SEM. Similar results were observed in independent duplicate experiments. Source data are provided as a Source Data file. f Transmigration assay of E0771 with recombinant galectin-3. Data were statistically analyzed with Holm-Sidak adjusted multiple t -test ( n = 3 biologically independent samples). The p -values are indicated in the graph. Data are presented as mean values ± SEM. Similar results were observed in independent duplicate experiments. Source data are provided as a Source Data file.

Article Snippet: For immunofluorescence staining, the fixed sections were washed with PBS containing 0.05% Tween-20 (Nacalai Tesque) (PBS-T) three times and then were incubated in blocking buffer (5% donkey serum, FUJIFILM Wako Pure Chemical Corporation) for 1 h at room temperature, and then incubated with anti-mouse galectin-3 primary antibody (CL8942AP, 1:2000, Cedarlane Laboratories, Burlington, Canada) diluted in PBS containing 1% bovine serum albumin (Nacalai Tesque) overnight at 4 °C.

Techniques: Marker, Expressing, Staining, Proliferation Assay, Transmigration Assay, Recombinant

Journal: The Journal of Biological Chemistry

Article Title: Distinct cargo-specific response landscapes underpin the complex and nuanced role of galectin–glycan interactions in clathrin-independent endocytosis

doi: 10.1074/jbc.RA118.001802

Figure Lengend Snippet: Galectin 3 is secreted and is bound to the surface and to MHCI of HeLa cells. A, endogenous galectin 3 is secreted into the medium and can be extracted from the cell surface. The galectin 3 content in complete medium, serum-free medium, and 48-h conditioned medium was measured using an ELISA. B, surface-bound galectin 3 was extracted from cells incubated with lactose-containing, but not sucrose-containing serum-free medium. C, HeLa cells were untreated or treated with 10 mm GlcNAc for 48 h before immunoprecipitation (IP) with antibodies to MHCI (w6/32). The gel on the left shows 2% lysate used as the input for immunoprecipitation, and the gel on the right shows the material pulled down with the beads. The blots were probed with antibodies to MHCI (HC110) and galectin 3, and representative Western blots are shown. D, quantification of MHCI-Gal3 co-immunoprecipitation Western blots plotting -fold change in galectin 3 normalized to MHCI pulled down for control and GlcNAc-treated conditions. At least three independent experiments were carried out with data expressed as mean ± S.D. (error bars). *, p < 0.05.

Article Snippet: Cells were transfected with GFP-tagged galectin 3 (Addgene) or GFP (Clontech).

Techniques: Enzyme-linked Immunosorbent Assay, Incubation, Immunoprecipitation, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Distinct cargo-specific response landscapes underpin the complex and nuanced role of galectin–glycan interactions in clathrin-independent endocytosis

doi: 10.1074/jbc.RA118.001802

Figure Lengend Snippet: MHCI and CD59 CIE have different responses to galectin 3, and CME is unaffected. Shown is antibody internalization for 30 min at 37 °C in HeLa cells after a 48-h incubation with or without 10 mm GlcNAc and with or without galectin 3 KD for MHCI (A) and CD59 (B). Galectin 3 depletion resulted in a stimulation of CD59 CIE and an ablation of GlcNAc-driven changes in MHCI CIE. C, flow cytometric analysis of the clathrin-mediated endocytosis of fluorescently labeled transferrin taken up over 10 min at 37 °C in HeLa cells shows that CME is unaffected by altering glycan interactions. D, Western blot analysis indicates that siRNA effectively depleted cells of galectin 3. E and F, the addition of exogenous recombinant galectin 3 along with the antibody in an antibody internalization assay resulted in an increase in MHCI internalization (E) and a decrease in CD59 internalization (F). At least three independent experiments were carried out with data expressed as mean ± S.D. (error bars). *, p < 0.05.

Article Snippet: Cells were transfected with GFP-tagged galectin 3 (Addgene) or GFP (Clontech).

Techniques: Incubation, Labeling, Western Blot, Recombinant

Journal: The Journal of Biological Chemistry

Article Title: Distinct cargo-specific response landscapes underpin the complex and nuanced role of galectin–glycan interactions in clathrin-independent endocytosis

doi: 10.1074/jbc.RA118.001802

Figure Lengend Snippet: The extent of the galectin lattice modulates cell spreading. After the relevant incubations, transfections, and pretreatments, HeLa cells were allowed to spread on a coverglass and fixed after 2 or 4 h, and the area of cell spreading was measured for each cell. Disrupting the galectin lattice with lactose treatment (A) or by knocking down galectin 3 (C) leads to an inhibition in cell spreading (at 4 h). A similar increase in osmolarity using 100 mm sucrose or 50 mm NaCl did not have any effect on cell spreading (at 4 h) (B). Increasing galectin lattice strength by increasing glycan branching (using GlcNAc treatment) (at 2 h) (D) or by overexpressing galectin 3 (at 4 h) (E) leads to an increase in cell spreading, and disrupting these strengthened lattices by lactose treatment ablates these increases. F, representative images of cells for the various treatments in the cell-spreading assay. Scale bars, 20 μm. G, a secretion assay measuring GFP fluorescence in the conditioned medium after 48 h indicates that Gal3-GFP is secreted by transfected HeLa cells into the medium. At least three independent experiments were carried out with data in column graphs expressed as mean ± S.D. (error bars); scatter dot plot data are expressed as median with interquartile range; *, p < 0.05.

Article Snippet: Cells were transfected with GFP-tagged galectin 3 (Addgene) or GFP (Clontech).

Techniques: Transfection, Inhibition, Fluorescence

Journal: The Journal of Biological Chemistry

Article Title: Distinct cargo-specific response landscapes underpin the complex and nuanced role of galectin–glycan interactions in clathrin-independent endocytosis

doi: 10.1074/jbc.RA118.001802

Figure Lengend Snippet: A conceptual interaction landscape illustrates the unique response profiles observed for each cargo. The strength of glycan-mediated interactions is represented along the horizontal axis with GlcNAc treatment, galectin 3 overexpression and the addition of exogenous galectin 3 increasing the strength of glycan-mediated interactions and shifting the cells to the right of the landscape. By contrast, galectin 3 knockdown inhibits galectin 3–dependent glycan interactions and shifts the cells to the left of the landscape and lactose treatment, which inhibits all galectin–glycan interactions, shifts the cells to the extreme left of the landscape. Glycosylation-sensitive endocytosis is represented on the vertical axis. Response curves are depicted for CD59 (in green) and MHCI (in red). The landscape also has a macropinocytosis aspect (in dashed lines), which cell lines that possess the requisite machinery (Beas2b and HT1080) can move through. The potential starting points on this landscape are indicated for each cell line based on the experimental observations. For HeLa cells, increasing glycan-mediated interactions (moving to the right) by GlcNAc treatment or the addition of exogenous galectin 3 led to an increase in MHCI CIE and a decrease in CD59 CIE, whereas inhibiting all glycan-mediated interactions by lactose treatment (moving to the far left) led to an inhibition of CIE for CD59. Inhibition of complex glycoform synthesis by swainsonine treatment, which also leads to a dramatic inhibition of galectin–glycan interactions (moving to the far left), also led to an inhibition of CIE for CD59. A more moderate decrease in glycan-mediated interactions by galectin 3 KD led to an increase in CD59 CIE. For HT1080 cells (which perform a lot of macropinocytosis), the cells start on the macropinocytosis aspect of the landscape. Inhibiting glycan-mediated interactions by lactose treatment (shift to the left) led to an increase in macropinocytosis, whereas an increase in glycan-mediated interactions by GlcNAc treatment (shift to the right) led to a decrease in macropinocytosis. For Beas2b cells (which perform macropinocytosis to a lesser degree), the cells start at an intermediate point on the landscape. Inhibiting glycan-mediated interactions by lactose treatment (shift to the left) pushes the cells up their macropinocytosis curve and leads to an increase in CIE of both cargos. In contrast, an increase in glycan-mediated interactions by GlcNAc treatment (shift to the right) pushed both cargos up their endocytic curves and led to an increase in CIE of both cargos.

Article Snippet: Cells were transfected with GFP-tagged galectin 3 (Addgene) or GFP (Clontech).

Techniques: Over Expression, Inhibition