Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 inhibition reduced lung tissues damage induced by lung I/R (A) The Gal3 protein expression in lung tissue was assessed by western blotting, and the density of Gal3 (relative to β-actin) on western blotting was quantified (n = 3). (B–D) The expression of Gal3 in lung tissue, BALF and serum was detected by ELISA (n = 6). (E) Paraffin-embedded lung tissue sections were stained with hematoxylin and eosin (n = 3) at magnifications of 200×, scale bar 200 μm and 400X, scale bar 100 μm. (F) The extent of lung injury was assessed by scoring the F images (n = 6 section per group). (G) The W/D ratio of lung tissue (n = 6). (H) Ultrastructural changes were evaluated through transmission electron microscopy. Arrows indicate lamellar body and triangles mitochondria (n = 3). Scale bar, 1 μm. The data were mean ± SEM. One-way analysis of variance was used for data comparison, ns, not significant, ∗p < 0.05, was statistically significant.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay, Staining, Transmission Assay, Electron Microscopy, Comparison

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 inhibition attenuated the inflammatory response induced by lung I/R (A–C) BALF of left lung after lung I/R was collected, and the supernatant was extracted to detect the contents of TNF-α, IL-1β, and IL-10 in BALF. (D–F) After LIRI, the heart blood was taken, and the supernatant was collected by centrifugation after standing at low temperature for 3 h. The levels of TNF-α, IL-1β, and IL-10 in the serum were detected. (G–I) The lung tissue was subjected to standard treatment to detect the contents of TNF-α, IL-1β, and IL-10 in lung tissue. Data shown are mean ± SEM from n = 6 per group, ns, no statistical significance, ∗p < 0.05, was statistically significant.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Centrifugation

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Inhibition of Gal3 reduces the expression of M1-type macrophages induced by lung I/R (A) Western blot of IL-1β, IL-6, TNF-α, and CCL5 in lung tissues. (B–E) Density analysis of IL-1β, IL-6, TNF-α, and CCL5 relative to β-actin in part A. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05). (F and G) Lung tissues were stained with macrophages marker (F4/80, green), anti-IL-1β (red), and anti-IL-6 antibodies (red). Nuclei were stained with DAPI (blue). Magnification, 400×, scale bar, 100 μm.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Expressing, Western Blot, Staining, Marker

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Inhibition of Gal3 increases the expression of M2 type macrophages induced by lung I/R (A) Western blot of Arg1, IL-10, Retnla, and Chil3 in lung tissues. (B–E) Density analysis of Arg1, IL-10, Retnla, and Chil3 relative to β-actin in part A. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05). (F and G) Lung tissues were stained with macrophages marker (F4/80, green), anti-Arg1 (red), and anti-IL-10 antibodies (red). Nuclei were stained with DAPI (blue). Magnification, 400×, scale bar, 100 μm.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Expressing, Western Blot, Staining, Marker

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 inhibition reduces macrophage polarization toward M1-type in OGD/R vitro model (A) Western blot of Gal3 in lung tissues. (B) Density analysis of Gal3 relative to β-actin in part A. (C) Cell viability was measured by CCK-8 assay (n = 6). (D) Western blot of IL-1β, IL-6, TNF-α, and CCL5 in RAW264.7 cells. (E–H) Densitometry of western blots in part D. Levels were standardized uniformly with β-actin. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05). (I and J) Immunofluorescence staining showed macrophages marker (F4/80, green), IL-1β (red), and IL-6 (red) in RAW264.7 cells. Nuclei were stained with DAPI (blue). Magnification, 200×, scale bar, 200 μm.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Western Blot, CCK-8 Assay, Immunofluorescence, Staining, Marker

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 inhibition promotes the polarization of macrophages toward the M2-type in OGD/R vitro model (A) Western blot of Arg1, IL-10, Retnla, and Chil3 in RAW264.7 cells. (B–E) Densitometry of western blots in part A. Levels were standardized uniformly with β-actin. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05). (F and G) Immunofluorescence staining showed macrophages marker (F4/80, green), Arg1 (red), and IL-10 (red) in RAW264.7 cells. Nuclei were stained with DAPI (blue). Magnification, 200×, scale bar, 200 μm.

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Inhibition, Western Blot, Immunofluorescence, Staining, Marker

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet: Gal3 and TREM2 colocalized in macrophages and Gal3 inhibited the expression of TREM2 in LIRI (A) Macrophage markers (F4/80, green), anti-TREM2 antibody (red), and anti-Gal3 antibody (pink) were used for immunofluorescence co-localization staining of lung tissue. Magnification, 400×, scale bar, 200 μm. (B) Enlarge the dotted box in part A. F4/80 (green), TREM2 (red), and Gal3 (pink) co-localization qualitative analysis. White dots to white triangles area (white dotted line) were analyzed with line intensity scans at higher magnification. Measurement of the intensity values demonstrated co-localization in those regions. Magnification, 2000×, scale bar, 50 μm. (C) Western blotting and (E) RT-qPCR expression of TREM2 in lung tissue. (D) Western blotting and (F) RT-qPCR expression of TREM2 in RAW264.7 cells. (n = 3, mean ± SEM, ns, no statistical significance, ∗p < 0.05).

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Expressing, Immunofluorescence, Staining, Western Blot, Quantitative RT-PCR

Journal: iScience

Article Title: Galectin-3 as TREM2 upstream factor contributes to lung ischemia-reperfusion injury by regulating macrophage polarization

doi: 10.1016/j.isci.2023.107496

Figure Lengend Snippet:

Article Snippet: Gal3 ELISA kit , CUSABIO , Cat #CSB-E14296m.

Techniques: Recombinant, Modification, Saline, Protease Inhibitor, Enzyme-linked Immunosorbent Assay

Journal: Biomolecules

Article Title: Exploration into Galectin-3 Driven Endocytosis and Lattices

doi: 10.3390/biom14091169

Figure Lengend Snippet: Gal3-based duality in β 1 integrin dynamics. ( A ) Schematic representation of the molecular organization of Gal3 where the C-terminal carbohydrate recognition domain (CRD) and the N-terminal oligomerization domain are indicated. ( B ) Left: Representative region of interest (leading edge) of a 2D STORM image of an RPE-1 cell showing surface-bound Gal3 ( top ) and the corresponding clusters obtained after segmentation ( bottom ). Right : The occurrence of each type of Gal3 cluster is shown in function of their surface area (×10 3 nm 2 ). Means ± SEM, one-way ANOVA; ns = p > 0.05, **** p < 0.0001. Scale bar = 5 μm. ( C ) The percentage of Gal3 molecules is shown in function of the different Gal3 cluster populations. Means ± SEM, one-way ANOVA; ns = p > 0.05, * p < 0.05, ** p < 0.002, **** p < 0.0001. ( D ) 2D STORM image of Gal3 and anti-β 1 integrin antibodies on RPE-1 cells (4 °C co-binding). A zoom of the leading edge is shown to illustrate the extensive level of colocalization between Gal3 and β 1 integrin. Gal3 clusters with variable size and shape are detected (yellow arrowheads in the zoom). Scale bars = 5 μm. (E) Quantification of the probability of proximity (colocalization) between Gal3 and β 1 integrin for each class of Gal3 clusters, compared to random non-clustered Gal3. Means ± SEM, one-way ANOVA; ns = p > 0.05, **** p < 0.0001. ( F ) Top: Anti-β 1 integrin antibody uptake assay in RPE-1 cells. Internalized antibody is immunolabeled, imaged by confocal microscopy, and quantified. Bottom: Scheme of the protocol detailing the use of the Gal3 inhibitor I3 (10 μM) in acute versus prolonged incubation conditions. ( G ) Anti-β 1 integrin antibody uptake assay as in ( F ). Note the shift from both peripheral (green arrowheads) and perinuclear (red arrowheads) distribution of internalized β 1 integrin in control cells to exclusive perinuclear localization in the prolonged incubation condition. ( H ) Transferrin (Tf) internalization is very little affected by I3. In ( G ) and ( H ): Dashed lines indicate the contour of individual cells. Scale bars = 10 μm. Nuclei in blue (DAPI). Quantification of fluorescence intensities as means ± SEM, one-way ANOVA; ns = p > 0.05, ** p < 0.002, **** p < 0.0001.

Article Snippet: Reagents: Anti-β 1 mAb13 antibody (BD Bioscience, Le Pont de Claix, France; Ref. 552828), anti-Gal3 antibody (Fu-Tong Liu, UC Davis, CA, USA), Cy3-labeled anti-β 1 integrin mAb13 antibody, anti-EEA1 antibody (Biorbyt, Durham, US; Ref. 11606), Cy3-labeled donkey anti-rat antibody (Beckman Coulter, Roissy CDG, France; Ref. 712-166-153), Cy3-labeled anti-goat antibody, fluorescein-isothiocyanate dextran 70 kDa (Sigma Merck, Saint Quentin Fallavier, France; Ref. 46945), Gal3 inhibitor compound I3 (1,1′-sulfanediyl-bis-{3-deoxy-3-[4-(butylaminocarbonyl)-1H-1,2,3-triazol-1-yl]-β-D galactopyranoside} [ , ], Genz-123346 (Sigma Merck, Saint Quentin Fallavier, France; Ref. 5382850001), β-D-lactose (Sigma Merck, Saint Quentin Fallavier, France; Ref. L3750), NHS-Cy3 (GE, Buc, France; PA23001), NHS-CF680 (Sigma Merck, Saint Quentin Fallavier, France; SCJ4600055), transferrin-Alexa546 (Tf-A546) (Invitrogen, Saint Aubin, France; Ref. T23364), AlexaFluor488-labeled recombinant purified Gal3, Cy3-labeled recombinant purified Gal3, AlexaFluor647-labeled recombinant purified Gal3.

Techniques: Binding Assay, Immunolabeling, Confocal Microscopy, Incubation, Control, Fluorescence

Journal: Biomolecules

Article Title: Exploration into Galectin-3 Driven Endocytosis and Lattices

doi: 10.3390/biom14091169

Figure Lengend Snippet: GSL-based duality in β 1 integrin dynamics. ( A ) Simplified schematic representation of the early steps of GSL synthesis. The reaction inhibited by Genz-123346 is indicated. ( B ) Analysis of cellular levels of the indicated GSL in function of incubation time with Genz-123346. Note that the most important drop occurs up to day 3. Means ± SEM, unpaired t -test; ns = p > 0.05, ** p < 0.002, **** p < 0.0001. ( C ) Scheme of experimental procedure detailing how GSL inhibition has been set up either in acute (3 days) or prolonged (5 days) incubation conditions, prior to cargo protein internalization for 10 min. ( D ) Anti-β 1 integrin antibody uptake assay as in ( C ). Note that β 1 integrin uptake is inhibited upon acute Genz-123346 treatment and increased upon prolonged treatment. In the latter condition, the intracellular accumulation of β 1 integrin is massively perinuclear (red arrowheads), compared to control cells where peripheral localizations are also observed (green arrowheads). Means ± SEM, unpaired t -test; **** p < 0.0001. ( E ) Transferrin (Tf) internalization (10 min) is only mildly affected in all conditions. Means ± SEM, unpaired t -test; ns = p > 0.05, ** p < 0.002. ( F ) Internalization of exogenous Gal3 (10 min). Similar to β 1 integrin, Gal3 endocytosis is significantly inhibited upon acute Genz-123346 treatment, and increased with perinuclear accumulation upon prolonged treatment (red arrowheads). Means ± SEM, unpaired t -test; **** p < 0.0001. In ( D – F ): Yellow dashed lines indicate contours of cells; scale bars = 10 μm, nuclei in blue (DAPI).

Article Snippet: Reagents: Anti-β 1 mAb13 antibody (BD Bioscience, Le Pont de Claix, France; Ref. 552828), anti-Gal3 antibody (Fu-Tong Liu, UC Davis, CA, USA), Cy3-labeled anti-β 1 integrin mAb13 antibody, anti-EEA1 antibody (Biorbyt, Durham, US; Ref. 11606), Cy3-labeled donkey anti-rat antibody (Beckman Coulter, Roissy CDG, France; Ref. 712-166-153), Cy3-labeled anti-goat antibody, fluorescein-isothiocyanate dextran 70 kDa (Sigma Merck, Saint Quentin Fallavier, France; Ref. 46945), Gal3 inhibitor compound I3 (1,1′-sulfanediyl-bis-{3-deoxy-3-[4-(butylaminocarbonyl)-1H-1,2,3-triazol-1-yl]-β-D galactopyranoside} [ , ], Genz-123346 (Sigma Merck, Saint Quentin Fallavier, France; Ref. 5382850001), β-D-lactose (Sigma Merck, Saint Quentin Fallavier, France; Ref. L3750), NHS-Cy3 (GE, Buc, France; PA23001), NHS-CF680 (Sigma Merck, Saint Quentin Fallavier, France; SCJ4600055), transferrin-Alexa546 (Tf-A546) (Invitrogen, Saint Aubin, France; Ref. T23364), AlexaFluor488-labeled recombinant purified Gal3, Cy3-labeled recombinant purified Gal3, AlexaFluor647-labeled recombinant purified Gal3.

Techniques: Incubation, Inhibition, Control

Journal: Biomolecules

Article Title: Exploration into Galectin-3 Driven Endocytosis and Lattices

doi: 10.3390/biom14091169

Figure Lengend Snippet: Characterization of sites of perinuclear β 1 integrin accumulation. ( A , B ) Anti-β 1 integrin or ( C ) Gal3 uptake assay (10 min) under acute or prolonged I3 ( A ) or Genz-123346 ( B , C ) treatment followed by immunolabeling for EEA1. The colocalization of β 1 integrin ( A , B ) or Gal3 ( C ) with EEA1 as well as the fluorescent intensity of EEA1 signal were quantified ( right ). Note the increased colocalization of internalized β 1 integrin ( A , B ) or Gal3 ( C ) with EEA1 and increased EEA1 signal intensity, notably in the prolonged treatment conditions. Means ± SEM, one-way ANOVA ( A , B ), or unpaired t -test ( C ); **** p < 0.0001. Yellow dashed lines indicate contours of cells. Scale bars = 10 μm, nuclei in blue (DAPI).

Article Snippet: Reagents: Anti-β 1 mAb13 antibody (BD Bioscience, Le Pont de Claix, France; Ref. 552828), anti-Gal3 antibody (Fu-Tong Liu, UC Davis, CA, USA), Cy3-labeled anti-β 1 integrin mAb13 antibody, anti-EEA1 antibody (Biorbyt, Durham, US; Ref. 11606), Cy3-labeled donkey anti-rat antibody (Beckman Coulter, Roissy CDG, France; Ref. 712-166-153), Cy3-labeled anti-goat antibody, fluorescein-isothiocyanate dextran 70 kDa (Sigma Merck, Saint Quentin Fallavier, France; Ref. 46945), Gal3 inhibitor compound I3 (1,1′-sulfanediyl-bis-{3-deoxy-3-[4-(butylaminocarbonyl)-1H-1,2,3-triazol-1-yl]-β-D galactopyranoside} [ , ], Genz-123346 (Sigma Merck, Saint Quentin Fallavier, France; Ref. 5382850001), β-D-lactose (Sigma Merck, Saint Quentin Fallavier, France; Ref. L3750), NHS-Cy3 (GE, Buc, France; PA23001), NHS-CF680 (Sigma Merck, Saint Quentin Fallavier, France; SCJ4600055), transferrin-Alexa546 (Tf-A546) (Invitrogen, Saint Aubin, France; Ref. T23364), AlexaFluor488-labeled recombinant purified Gal3, Cy3-labeled recombinant purified Gal3, AlexaFluor647-labeled recombinant purified Gal3.

Techniques: Immunolabeling

Journal: Biomolecules

Article Title: Exploration into Galectin-3 Driven Endocytosis and Lattices

doi: 10.3390/biom14091169

Figure Lengend Snippet: Exogenous Gal3 and dextran 70K uptake upon prolonged GSL depletion. After prolonged (5 days) treatment with Genz-123346, RPE-1 cells were continuously co-incubated (10 min) with exogenous Gal3 and dextran 70K. Note the increased perinuclear accumulation of Gal3 and its increased overlap with dextran 70K under these conditions. Means ± SEM, unpaired t -test; **** p < 0.0001. Yellow dashed lines indicate contours of cells. Scale bars = 10 μm, nuclei in blue (DAPI).

Article Snippet: Reagents: Anti-β 1 mAb13 antibody (BD Bioscience, Le Pont de Claix, France; Ref. 552828), anti-Gal3 antibody (Fu-Tong Liu, UC Davis, CA, USA), Cy3-labeled anti-β 1 integrin mAb13 antibody, anti-EEA1 antibody (Biorbyt, Durham, US; Ref. 11606), Cy3-labeled donkey anti-rat antibody (Beckman Coulter, Roissy CDG, France; Ref. 712-166-153), Cy3-labeled anti-goat antibody, fluorescein-isothiocyanate dextran 70 kDa (Sigma Merck, Saint Quentin Fallavier, France; Ref. 46945), Gal3 inhibitor compound I3 (1,1′-sulfanediyl-bis-{3-deoxy-3-[4-(butylaminocarbonyl)-1H-1,2,3-triazol-1-yl]-β-D galactopyranoside} [ , ], Genz-123346 (Sigma Merck, Saint Quentin Fallavier, France; Ref. 5382850001), β-D-lactose (Sigma Merck, Saint Quentin Fallavier, France; Ref. L3750), NHS-Cy3 (GE, Buc, France; PA23001), NHS-CF680 (Sigma Merck, Saint Quentin Fallavier, France; SCJ4600055), transferrin-Alexa546 (Tf-A546) (Invitrogen, Saint Aubin, France; Ref. T23364), AlexaFluor488-labeled recombinant purified Gal3, Cy3-labeled recombinant purified Gal3, AlexaFluor647-labeled recombinant purified Gal3.

Techniques: Incubation

Journal: Biomolecules

Article Title: Exploration into Galectin-3 Driven Endocytosis and Lattices

doi: 10.3390/biom14091169

Figure Lengend Snippet: Role of clathrin in endocytic uptake under prolonged treatment conditions. ( A – C ) Uptake assays (10 min) of anti-β 1 integrin antibodies ( A , B ) or Gal3 ( C ) upon prolonged I3 ( A ) or Genz-123346 ( B , C ) treatment. When indicated (siCHC), clathrin heavy chain was depleted ( right images). The perinuclear accumulation of β 1 integrin ( A , B ) or that of Gal3 ( C ) as observed in the prolonged treatment conditions (red or white arrowheads) is strongly inhibited upon clathrin depletion. Means ± SEM, one-way ANOVA; **** p < 0.0001. Yellow dashed lines indicate contours of cells. Scale bars = 10 μm, nuclei in blue (DAPI).

Article Snippet: Reagents: Anti-β 1 mAb13 antibody (BD Bioscience, Le Pont de Claix, France; Ref. 552828), anti-Gal3 antibody (Fu-Tong Liu, UC Davis, CA, USA), Cy3-labeled anti-β 1 integrin mAb13 antibody, anti-EEA1 antibody (Biorbyt, Durham, US; Ref. 11606), Cy3-labeled donkey anti-rat antibody (Beckman Coulter, Roissy CDG, France; Ref. 712-166-153), Cy3-labeled anti-goat antibody, fluorescein-isothiocyanate dextran 70 kDa (Sigma Merck, Saint Quentin Fallavier, France; Ref. 46945), Gal3 inhibitor compound I3 (1,1′-sulfanediyl-bis-{3-deoxy-3-[4-(butylaminocarbonyl)-1H-1,2,3-triazol-1-yl]-β-D galactopyranoside} [ , ], Genz-123346 (Sigma Merck, Saint Quentin Fallavier, France; Ref. 5382850001), β-D-lactose (Sigma Merck, Saint Quentin Fallavier, France; Ref. L3750), NHS-Cy3 (GE, Buc, France; PA23001), NHS-CF680 (Sigma Merck, Saint Quentin Fallavier, France; SCJ4600055), transferrin-Alexa546 (Tf-A546) (Invitrogen, Saint Aubin, France; Ref. T23364), AlexaFluor488-labeled recombinant purified Gal3, Cy3-labeled recombinant purified Gal3, AlexaFluor647-labeled recombinant purified Gal3.

Techniques:

Journal: Biomolecules

Article Title: Exploration into Galectin-3 Driven Endocytosis and Lattices

doi: 10.3390/biom14091169

Figure Lengend Snippet: Continuum model between lattices and GL-Lect driven endocytosis. ( A ) Unperturbed condition. A glycoprotein cargo, here α 5 β 1 integrin, is either recruited into galectin lattices ( left , underlined in red) or internalized by GL-Lect driven endocytosis ( right , underlined in blue). ( B ) Acute treatment conditions. Since tubular endocytic pits for GL-Lect driven endocytosis are built de novo, acute interference with Gal3 activity or GSL expression prevents their formation. In contrast, preassembled galectin lattices resist under these conditions. ( C ) Prolonged treatment conditions. Even galectin lattices are disassembled. With GL-Lect driven endocytosis being inhibited, α 5 β 1 integrin is now internalized by alternative endocytic pathways, i.e., clathrin-mediated endocytosis and macropinocytosis.

Article Snippet: Reagents: Anti-β 1 mAb13 antibody (BD Bioscience, Le Pont de Claix, France; Ref. 552828), anti-Gal3 antibody (Fu-Tong Liu, UC Davis, CA, USA), Cy3-labeled anti-β 1 integrin mAb13 antibody, anti-EEA1 antibody (Biorbyt, Durham, US; Ref. 11606), Cy3-labeled donkey anti-rat antibody (Beckman Coulter, Roissy CDG, France; Ref. 712-166-153), Cy3-labeled anti-goat antibody, fluorescein-isothiocyanate dextran 70 kDa (Sigma Merck, Saint Quentin Fallavier, France; Ref. 46945), Gal3 inhibitor compound I3 (1,1′-sulfanediyl-bis-{3-deoxy-3-[4-(butylaminocarbonyl)-1H-1,2,3-triazol-1-yl]-β-D galactopyranoside} [ , ], Genz-123346 (Sigma Merck, Saint Quentin Fallavier, France; Ref. 5382850001), β-D-lactose (Sigma Merck, Saint Quentin Fallavier, France; Ref. L3750), NHS-Cy3 (GE, Buc, France; PA23001), NHS-CF680 (Sigma Merck, Saint Quentin Fallavier, France; SCJ4600055), transferrin-Alexa546 (Tf-A546) (Invitrogen, Saint Aubin, France; Ref. T23364), AlexaFluor488-labeled recombinant purified Gal3, Cy3-labeled recombinant purified Gal3, AlexaFluor647-labeled recombinant purified Gal3.

Techniques: Activity Assay, Expressing