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