Journal: Breast Cancer Research : BCR

Article Title: Secreted LGALS3BP facilitates distant metastasis of breast cancer

doi: 10.1186/s13058-024-01958-8

Figure Lengend Snippet: Secretion of LGAL3BP increased from the MCF7/TAMR and T47D/TR cells. ( A ) Brief experimental scheme for Fig. 2. ( B ) Whole cell lysates obtained from the MCF7/TAMR and T47D/TR cells and their parental cells were subjected to western blotting (upper). The proteins in the culture supernatants were TCA precipitated and immunoblotted with anti-LGALS3BP antibodies (lower). Band intensity was quantified and normalized to the intensity of the corresponding β-actin band. ( C ) The amounts of LGALS3BP in the culture supernatants were measured by ELISA. ( D ) Expression of LGALS3BP was visualized by green immunofluorescence. The nuclei were stained with DAPI. Scale bars: 20 μm. Data presented as mean ± SD ( n = 3). *, P < 0.05; **, P < 0.01, and ***, P < 0.001

Article Snippet: The recombinant LGALS3BP protein (2226-GAB) was purchased from R&D Systems.

Techniques: Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Immunofluorescence, Staining

Journal: Breast Cancer Research : BCR

Article Title: Secreted LGALS3BP facilitates distant metastasis of breast cancer

doi: 10.1186/s13058-024-01958-8

Figure Lengend Snippet: LGALS3BP enhances cell adhesion of MCF7/TAMR cells and tube formation of HUVEC. ( A ) Attachment of the MCF7/TAMR, shLG, and their matched control cells was evaluated by staining with crystal violet 2 h after seeding (upper). Or MCF7/TAMR-8 cells were pre-treated with 1 µg/mL of anti-LGALS3BP antibodies or IgG control for 10 min before the adhesion assay was performed (lower). Adherent cells were quantified by dissolving them in MeOH before the absorbance at 570 nm was detected. Scale bars = 500 μm. ( B ) HUVECs were cultured in conditioned media obtained from MCF7/TAMR, shLG, and their matched control cells (upper). Or HUVECs were cultured in MCF7/TAMR-8 conditioned media treated with 1 µg/mL of anti-LGALS3BP antibodies or IgG control (lower). Tube formation was quantified by counting the number of meshes formed. Scale bars = 100 μm. ( C ) HUVECs were cultured in serum-free or conditioned media obtained from shLG #1 cells and treated with recombinant LGALS3BP protein as indicated. Tube formation was quantified by counting the number of meshes formed. Scale bars = 100 μm. rLG, recombinant LGALS3BP ( D ) HUVECs were treated with recombinant LGALS3BP protein as indicated. The cell growth was determined using MTT assay. Data presented as mean ± SD. *, P < 0.05, **, P < 0.01, and ***, P < 0.001 ( n = 3 for A and D and n = 4 for B and C)

Article Snippet: The recombinant LGALS3BP protein (2226-GAB) was purchased from R&D Systems.

Techniques: Control, Staining, Cell Adhesion Assay, Cell Culture, Recombinant, MTT Assay

Journal: Breast Cancer Research : BCR

Article Title: Secreted LGALS3BP facilitates distant metastasis of breast cancer

doi: 10.1186/s13058-024-01958-8

Figure Lengend Snippet: Depletion of LGALS3BP suppresses pulmonary metastasis of the MCF7/TAMR-8 in xenograft experiments. ( A ) MCF7/S0.5 or TAMR-8 cells were inoculated into the fourth mammary fat pads of female athymic nude mice. When the tumor volume reached approximately 50 mm 3 , the tumor diameter was measured with a caliper ( n = 9–10). ( B ) Representative images of LGALS3BP immunofluorescent staining in the primary tumor sections. Scale bars = 20 μm. ( C ) The spontaneous lung metastasis developed from the MCF7/TAMR cells and their parental cell xenografts. Hematoxylin and eosin staining of lung sections are shown. Arrowheads indicate metastatic lesions. Scale bars = 800 μm for full images and 200 μm for inserts. The area of metastatic lesions in the lungs were quantified using ImageJ. Data presented as mean ± SD ( n = 9–10). *, P < 0.05. ( D ) MCF7/TAMR-8-shScramble or MCF7/TAMR-8-shLG cells were inoculated into the fourth mammary fat pads of female athymic nude mice. When the tumor volume reached approximately 50 mm 3 , the tumor diameter was measured with a caliper. ( n = 9) ( E ) Representative images of LGALS3BP immunofluorescent staining in the primary tumor sections. Scale bars = 20 μm. ( F ) The spontaneous lung metastasis developed from the shLG and its control cell xenograft. Hematoxylin and eosin staining of lung sections are shown. Arrowheads indicate metastatic lesions. Scale bars = 800 μm for full images and 200 μm for inserts. The area of the metastatic lesions in the lung was quantified using ImageJ. Data presented as mean ± SD ( n = 9). *, P < 0.05. ( G ) Immunohistochemistry of CD31 or fibronectin of shScrb and shLG primary tumors are shown. Scale bars = 100 μm. At least two images of 2 mm 3 in size from each primary tumor sections were quantified using ImageJ. Data presented as mean ± SD ( n = 7–8). *, P < 0.05

Article Snippet: The recombinant LGALS3BP protein (2226-GAB) was purchased from R&D Systems.

Techniques: Staining, Control, Immunohistochemistry

Journal: Breast Cancer Research : BCR

Article Title: Secreted LGALS3BP facilitates distant metastasis of breast cancer

doi: 10.1186/s13058-024-01958-8

Figure Lengend Snippet: Expression level of LGALS3BP is enhanced in the relapsed BC in clinic. ( A ) The expression level of LGALS3BP was evaluated by IHC in a total of 16 sets of primary and relapsed BC samples that obtained from patients with ER-positive BC. All patients received tamoxifen as adjuvant treatment . Representative images for immunohistochemical staining of LGALS3BP in the human breast cancer specimens. LGALS3BP staining intensity was scored by a board-certified pathologist. The comparison was made between primary tumor and matching relapsed tumor. Scale bars = 100 μm. * P < 0.05. ( B ) Kaplan-Meier analyses (log-rank tests) for relapse-free and distant-metastasis free survival of patients with ER-positive BC treated with tamoxifen were performed using KM plotter (GSE17705, GSE12093, GSE9195, GSE2990, and GSE45255) and GSE9893 dataset. High and low groups were defined based on maximal cut-off values

Article Snippet: The recombinant LGALS3BP protein (2226-GAB) was purchased from R&D Systems.

Techniques: Expressing, Adjuvant, Immunohistochemical staining, Staining, Comparison

Journal: Oncotarget

Article Title: Radiation-enhanced therapeutic targeting of galectin-1 enriched malignant stroma in triple negative breast cancer

doi: 10.18632/oncotarget.9490

Figure Lengend Snippet: ( A ) Spot images of human breast cancer (ductal carcinoma) and normal breast tissues 1 mm in diameter of women 27–40 years in age stained with galectin-1 antibody (Sigma) [Upper panel] and VEGF antibody (Santa Cruz Biotech.) [Lower panel]. ( B ) Quantitation of image scans (Aperioscope) demonstrating higher % of positive staining for both galectin-1 and VEGF-A in ductal carcinoma versus normal breast tissue with * p < 0.0001 and ** p < 0.05 respectively (mean ± SE; n = 3 samples/group). ( C ) 20X magnification of ductal carcinoma sections in (A) immunoprobed for galectin-1 demonstrating its stromal enrichment (Brown). Tumor cells stain blue (DAPI). Data were provided by the human protein atlas database. Scale bar = 100 μm.

Article Snippet: Slides were incubated in Galectin-1 antibody (R&D AF-1152) diluted at 1:50 for one hour at room temperature followed by incubation with polymer bound anti-goat secondary for 30 minutes (Vector Laboratories Immpress kit).

Techniques: Staining, Quantitation Assay

Journal: Oncotarget

Article Title: Radiation-enhanced therapeutic targeting of galectin-1 enriched malignant stroma in triple negative breast cancer

doi: 10.18632/oncotarget.9490

Figure Lengend Snippet: ( A ) Representative spot images of six types of tissues 1 mm in diameter as indicated were probed (Brown) for the expression of galectin-1 (Sigma) were compared to tumor tissue of ductal carcinoma stained for galectin-1. Data were provided by the human protein atlas database ( http://www.proteinatlas.org/ ). ( B ) Galectin-1 staining was quantitated by APERIO ® ImageScope and scored using the H-score approach that combines the pixel intensity with the percentage of tissue as described in the methods and graphically represented. ( N = 3, * p < 0.005 or ** p < 0.05, for tissues from the tumor compared to that from other organs).

Article Snippet: Slides were incubated in Galectin-1 antibody (R&D AF-1152) diluted at 1:50 for one hour at room temperature followed by incubation with polymer bound anti-goat secondary for 30 minutes (Vector Laboratories Immpress kit).

Techniques: Expressing, Staining

Journal: Oncotarget

Article Title: Radiation-enhanced therapeutic targeting of galectin-1 enriched malignant stroma in triple negative breast cancer

doi: 10.18632/oncotarget.9490

Figure Lengend Snippet: ( A ) Immunohistochemistry of six cases of TNBC matched with the corresponding normal breast tissues for galectin-1. The tissue sections were stained with anti-galectin-1 antibody (sigma) and counterstained with Hematoxylin and Eosin at 100X magnification. Scale bar = 25 μm ( B ) Galectin-1 staining in each histological section of benign and tumor tissues of the six TNBC patients was scored using the H-score approach as described in the methods and graphically represented. The percentage of tissue stained was quantitated by APERIO ® ImageScope ( N = 6, * p = 0.0002 for tumor tissue compared to the benign tissue). ( C ) Representative images of benign and primary tumor tissue probed for galectin-1. While the tumor cells stained weakly (blue/light brown), the tumor associated stroma stained strongly for galectin-1 (brown) in the tumor tissue. The myoepithelial cells/clusters and ductal/lobular units in the benign breast tissue also stained strongly for galectin-1 (black arrows) but the staining was restricted to distinct foci. Scale bar = 50 μm.

Article Snippet: Slides were incubated in Galectin-1 antibody (R&D AF-1152) diluted at 1:50 for one hour at room temperature followed by incubation with polymer bound anti-goat secondary for 30 minutes (Vector Laboratories Immpress kit).

Techniques: Immunohistochemistry, Staining

Journal: Oncotarget

Article Title: Radiation-enhanced therapeutic targeting of galectin-1 enriched malignant stroma in triple negative breast cancer

doi: 10.18632/oncotarget.9490

Figure Lengend Snippet: ( A ) Representative images of immunohistochemistry for galectin-1 in benign breast tissue (BT), tumor tissue (TT) and irradiated tumor tissue (IT) originating from orthotopic implants of TTA in mice 72 hours post-radiation exposure of 2 Gy. ( B ) Galectin-1 staining was quantitated by APERIO ® ImageScope and scored using the H-score approach as described in the methods and graphically represented. N = 3/group, * p < 0.0003 for TT vs IT, BT vs. IT and BT vs TT.

Article Snippet: Slides were incubated in Galectin-1 antibody (R&D AF-1152) diluted at 1:50 for one hour at room temperature followed by incubation with polymer bound anti-goat secondary for 30 minutes (Vector Laboratories Immpress kit).

Techniques: Immunohistochemistry, Irradiation, Staining

Journal: bioRxiv

Article Title: Expression and role of Galectin-3 in the postnatal development of the cerebellum

doi: 10.1101/364760

Figure Lengend Snippet: (A) Lgals3 mRNA expression relative to Rpl13a measured by RTqPCR in the cerebellum during development. E17: embryonic day 17. P0, P7, P14, and P21: postnatal day 0, 7, 14, and 21. (B) Lgals3 knockout (KO) mice present no defects in cerebellar morphology. Parasagittal cerebellar slices of wild-type (WT) and Lgals3 KO adult mice were stained using the nuclear marker Hoechst.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-CABP (1:1000; swant, Cat#300), rabbit polyclonal anti-CABP (1:1000; swant, Cat#9.03), mouse monoclonal anti-GFAP (1:500; millipore, Cat#MAB360), rabbit polyclonal anti-GLURδ1/2 (1:1000; millipore, Cat#AB2285), goat polyclonal anti-LGALS3 (1:200; R&D Systems, Cat#AF1197), mouse monoclonal anti-OLIG2 (1:500; millipore, Cat#MABN50), guinea pig polyclonal anti-VGLUT1 (1:5000; millipore, Cat#AB5905) and guinea pig polyclonal anti-VGLUT2 (1:5000; millipore, Cat#AB2251).

Techniques: Expressing, Knock-Out, Staining, Marker

Journal: bioRxiv

Article Title: Expression and role of Galectin-3 in the postnatal development of the cerebellum

doi: 10.1101/364760

Figure Lengend Snippet: (A and B) Immunostaining for the Purkinje cell marker CABP (green) and the presynaptic marker VGLUT2 (red) was performed on sections from Lgals3 WT and KO mice at P15 (A) and at P65 (B). (C and D) Immunostaining for the Purkinje cell marker CABP (green) and the presynaptic marker VGLUT1 (red) specific for mature parallel fiber synapses was performed on sections from Lgals3 WT and KO mice at P15 (C) and at P65 (D). Scale bar: 25μm. Data are representative of three independent experiments.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-CABP (1:1000; swant, Cat#300), rabbit polyclonal anti-CABP (1:1000; swant, Cat#9.03), mouse monoclonal anti-GFAP (1:500; millipore, Cat#MAB360), rabbit polyclonal anti-GLURδ1/2 (1:1000; millipore, Cat#AB2285), goat polyclonal anti-LGALS3 (1:200; R&D Systems, Cat#AF1197), mouse monoclonal anti-OLIG2 (1:500; millipore, Cat#MABN50), guinea pig polyclonal anti-VGLUT1 (1:5000; millipore, Cat#AB5905) and guinea pig polyclonal anti-VGLUT2 (1:5000; millipore, Cat#AB2251).

Techniques: Immunostaining, Marker

Journal: bioRxiv

Article Title: Expression and role of Galectin-3 in the postnatal development of the cerebellum

doi: 10.1101/364760

Figure Lengend Snippet: (A) Immunohistochemistry for LGALS3 show the specificity of the LGALS3 antibody. WT: wild-type mice; KO: knockout mice. Positive cells in the molecular layer are marked with a white arrowhead, scale bar 50μm. (B) Immunohistochemistry for the Purkinje cell marker CABP (green) and LGALS3 (red) on parasagittal cerebellar sections from P15 and P22 wild-type mice. Scale bar=100μm. Data are representative of three independent experiments. Legends: 4V, 4 th ventricle; chp, choroid plexus; pcg, pontine central gray and mv, medial vestibular nucleus.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-CABP (1:1000; swant, Cat#300), rabbit polyclonal anti-CABP (1:1000; swant, Cat#9.03), mouse monoclonal anti-GFAP (1:500; millipore, Cat#MAB360), rabbit polyclonal anti-GLURδ1/2 (1:1000; millipore, Cat#AB2285), goat polyclonal anti-LGALS3 (1:200; R&D Systems, Cat#AF1197), mouse monoclonal anti-OLIG2 (1:500; millipore, Cat#MABN50), guinea pig polyclonal anti-VGLUT1 (1:5000; millipore, Cat#AB5905) and guinea pig polyclonal anti-VGLUT2 (1:5000; millipore, Cat#AB2251).

Techniques: Immunohistochemistry, Knock-Out, Marker

Journal: bioRxiv

Article Title: Expression and role of Galectin-3 in the postnatal development of the cerebellum

doi: 10.1101/364760

Figure Lengend Snippet: (A) LGALS3 expression in oligodendrocytes in the cerebellar white matter. Co-immunostaining of parasagittal cerebellar slices using anti-LGALS3 (red), oligodendrocyte marker anti-OLIG-2 (green) and the nuclear marker Hoechst (blue). (B) LGALS3 expression in astrocytes in the cerebellar white matter. Co-immunostaining of parasagittal cerebellar slices using anti-LGALS3 (red), astrocyte marker anti-GFAP (green) and the nuclear marker Hoechst (blue). (C) LGALS3 expression in some microglial cells of the molecular layer. Immunostaining anti-LGALS3 (red) in parasagittal sections of mice CX 3 CR1 eGFP/eGFP , microglial CX 3 CR1-GFP (green) and the nuclear marker Hoechst (blue). Scale bar 50μm and 20μm for the magnification. Data are representative of three independent experiments. Legends: 4V, 4 th ventricle; GL, granular layer; ML, molecular layer and WM, white matter.

Article Snippet: The following primary antibodies were used: mouse monoclonal anti-CABP (1:1000; swant, Cat#300), rabbit polyclonal anti-CABP (1:1000; swant, Cat#9.03), mouse monoclonal anti-GFAP (1:500; millipore, Cat#MAB360), rabbit polyclonal anti-GLURδ1/2 (1:1000; millipore, Cat#AB2285), goat polyclonal anti-LGALS3 (1:200; R&D Systems, Cat#AF1197), mouse monoclonal anti-OLIG2 (1:500; millipore, Cat#MABN50), guinea pig polyclonal anti-VGLUT1 (1:5000; millipore, Cat#AB5905) and guinea pig polyclonal anti-VGLUT2 (1:5000; millipore, Cat#AB2251).

Techniques: Expressing, Immunostaining, Marker