Journal: Cancers

Article Title: Quercetin Induces Anticancer Activity by Upregulating Pro-NAG-1/GDF15 in Differentiated Thyroid Cancer Cells

doi: 10.3390/cancers13123022

Figure Lengend Snippet: Antibody array using human thyroid tissues. ( A ) Antibody array (RayBio ® C-Series Human Cancer Discovery Antibody Array 3, RayBiotech) showed that 30 cytokines related to cancer biology were differentially expressed between thyroid normal and tumor tissues. One pair of PTC samples was used to analyze the expression of these cytokines. Four different proteins (Galectin-3, NAG-1/GDF15, TIMP-1, and osteoprotegerin) were selected as biomarker candidates for the diagnosis of thyroid cancer (rectangle). The right graph represents the intensity of each protein. ( B ) Western blot was performed using three pairs of PTC samples to confirm the data of antibody array. The protein expression of galectin-3, mature NAG-1, pro-NAG-1, TIMP-1, and OPG was examined using thyroid tissue samples. The right graph is from the average quantification of protein expression from three patients. N, thyroid normal tissue; T, thyroid tumor tissue. The number of patients is indicated (see for details). Uncropped versions of blots presented in .

Article Snippet: The primary antibodies used were anti-NAG-1 (specific to both pro-NAG-1 and mature NAG-1) [ ], anti-galectin-3 (sc-32790; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-osteoprotegerin (OPG; sc-390518; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-TIMP-1(sc-6832; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-GAPDH (sc-47724; Santa Cruz Biotechnology, Santa Cruz, CA, USA), and anti-β-actin (sc-47778; Santa Cruz Biotechnology, Santa Cruz, CA, USA).

Techniques: Ab Array, Expressing, Biomarker Assay, Western Blot

Journal: STAR protocols

Article Title: Protocol for detecting lysosome quantity and membrane permeability in acute myeloid leukemia cell lines.

doi: 10.1016/j.xpro.2024.103309

Figure Lengend Snippet: Figure 3. Validation of LGALS3-EGFP expression in MV4-11 cells (A) The constructed MV4-11 cells expressing LGALS3-EGFP are observed under an inverted fluorescence microscope by using a green fluorescence filter set. In this assay, the wild-type MV4-11 cells are employed as a negative control. (B) The validation is conducted by measuring the fluorescence by using a 488 nm laser with 525/40 (Green) filters using DxFLEX (Beckman Coulter). In this assay, the wild-type MV4-11 cells are employed as a negative control.

Article Snippet: Alternatives: The LGALS3 plasmid used is also available via Addgene.

Techniques: Biomarker Discovery, Expressing, Construct, Microscopy, Negative Control

Journal: Journal of translational medicine

Article Title: LGALS3BP antibody-drug conjugate enhances tumor-infiltrating lymphocytes and synergizes with immunotherapy to restrain neuroblastoma growth.

doi: 10.1186/s12967-025-06434-1

Figure Lengend Snippet: Fig. 1 EVs-associated LGALS3BP levels correlate with metastatatic lesions and increase accordingly over time in a pseudometastatic model of human neuroblastoma. (A) Representative scheme of pseudometastatic model: human neuroblastoma SKNAS cells were tail-vein injected in NSG mice and blood was collected after 7 (n = 2), 14 (n = 3), 21 (n = 3), and 28 (n = 2) days. (B) EVs-associated LGALS3BP levels detected in ELISA (pg/mL) per µg of EVs isolated from mouse serum after 7 (n = 2), 14 (n = 3), 21 (n = 3), or 28 (n = 2) days. Tumor free mice (n = 2) were considered as controls. (C) Immunohis tochemistry staining with hematoxylin/eosin for metastatic lesions and LGALS3BP levels in metastatic lesions over time. Scale bar: 250 μm. Number of metastases detected in (D) liver, (E) lung, and (F) kidney of NSG mice after 7 (n = 2), 14 (n = 3), 21 (n = 3), and 28 (n = 2) days

Article Snippet: For the evaluation of serum and EVs-associated human LGALS3BP levels in human neuroblastoma pseudometastatic experiments, human LGALS3BP ELISA Kit was performed according to manufacturer’s instructions (#KE00155; Proteintech, USA).

Techniques: Injection, Enzyme-linked Immunosorbent Assay, Isolation, Staining

Journal: Journal of translational medicine

Article Title: LGALS3BP antibody-drug conjugate enhances tumor-infiltrating lymphocytes and synergizes with immunotherapy to restrain neuroblastoma growth.

doi: 10.1186/s12967-025-06434-1

Figure Lengend Snippet: Fig. 2 Circulating and EVs-associated LGALS3BP could function as early detection biomarker of neuroblastoma lesions in a human neuroblastoma pseu dometastatic model. (A) EVs-associated LGALS3BP levels detected in ELISA (pg/mL) per µg of EVs isolated from mouse serum collected 14 days after NSG mice (n = 6) were tail-vein injected with human neuroblastoma SKNAS cells. Tumor free mice (n = 2) were considered as controls. (B) Representative NTA profile of EVs isolated from mouse serum after 14 days from tail-vein injection of SKNAS cells. (C) Circulating LGALS3BP levels detected in ELISA (pg/mL) in mouse serum (diluted 1:4) after 14 days (n = 6). Tumor free mice (n = 2) were considered as controls. Number of metastases detected in (D) liver, (E) lung, and (F) kidney of NSG at 14 days post-injection

Article Snippet: For the evaluation of serum and EVs-associated human LGALS3BP levels in human neuroblastoma pseudometastatic experiments, human LGALS3BP ELISA Kit was performed according to manufacturer’s instructions (#KE00155; Proteintech, USA).

Techniques: Biomarker Discovery, Enzyme-linked Immunosorbent Assay, Isolation, Injection

Journal: Journal of translational medicine

Article Title: LGALS3BP antibody-drug conjugate enhances tumor-infiltrating lymphocytes and synergizes with immunotherapy to restrain neuroblastoma growth.

doi: 10.1186/s12967-025-06434-1

Figure Lengend Snippet: Fig. 3 Human LGALS3BP is overexpressed and secreted by engineered murine neuroblastoma NXS2 hLGALS3BP cells. (A) Histograms showing transcrip tional levels of human (hLGALS3BP) and murine (mLGALS3BP) LGALS3BP in NXS2 CTRL and hLGALS3BP cells normalized to housekeeping gene (β-actin). Values refer to two independent biological replicates. (B) Western blotting images showing intracellular protein levels of hLGALS3BP in NXS2 CTRL and hLGALS3BP cells. Equal amounts of protein were loaded for each sample. β-actin was used as loading control. Images are representative of two indepen dent biological replicates. (C) Secreted levels (ng/mL) of hLGALS3BP and mLGALS3BP assessed by ELISA assay in NXS2 CTRL and hLGALS3BP cells. Values refer to two independent biological replicates. (D) Table showing secreted LGALS3BP expression level in a panel of human cancer cell lines. (E) Confocal imaging of NXS2 CTRL and hLGALS3BP cells showing LGALS3BP (green) and nuclei (Hoechst). Scale bar: 10 μm. (F) Immunoblot showing EVs-associated hLGALS3BP and positive/negative exosomal markers expression in whole cell lysates (WCL) or EVs isolated from NXS2 CTRL or hLGALS3BP. Equal amount of protein were loaded for each sample.

Article Snippet: For the evaluation of serum and EVs-associated human LGALS3BP levels in human neuroblastoma pseudometastatic experiments, human LGALS3BP ELISA Kit was performed according to manufacturer’s instructions (#KE00155; Proteintech, USA).

Techniques: Western Blot, Control, Enzyme-linked Immunosorbent Assay, Expressing, Imaging, Isolation

Journal: Journal of translational medicine

Article Title: LGALS3BP antibody-drug conjugate enhances tumor-infiltrating lymphocytes and synergizes with immunotherapy to restrain neuroblastoma growth.

doi: 10.1186/s12967-025-06434-1

Figure Lengend Snippet: Fig. 5 Targeted and effective anti-tumor activity of anti-LGALS3BP ADC 1959-sss/DM4 in murine syngeneic neuroblastoma model. (A) Tumor growth expressed as tumor volume (mm3) from first treatment of NXS2 CTRL or hLGALS3BP allografts in syngeneic A/J mice treated or not with ADC 1959-sss/ DM4 (10 mg/kg; twice weekly; three total i.v. injections). Arrows indicate treatments administration. 4 or 5 mice per group were considered. (B) Kaplan- Meier curve showing probability of survival in percentage of mice bearing NXS2 CTRL or hLGALS3BP allografts treated or not with ADC 1959-sss/DM4 from first treatment. Log-rank (Mantel-Cox) test: ** p < 0.01. (C) TILs flow cytometric analysis of PBS (n = 4), 1959-sss (n = 2) and 1959-sss/DM4 (n = 4) treated tumors at 7 days after starting treatments, showing percentage of activated CD69 + within the total population of CD3+ T cells (left panel); percentage of activated CD8+ CD69+ within the total population of CD3+ T cells (middle panel). In the right panel is shown the percentage of CD69+ Granzyme B+ over cytotoxic CD8+ T cells. Differences were compared using unpaired T test, and considering significant p < 0.05, as indicated with an asterisk (* p < 0.05, ** p < 0.01, *** p < 0.001)

Article Snippet: For the evaluation of serum and EVs-associated human LGALS3BP levels in human neuroblastoma pseudometastatic experiments, human LGALS3BP ELISA Kit was performed according to manufacturer’s instructions (#KE00155; Proteintech, USA).

Techniques: Activity Assay

Journal: Journal of translational medicine

Article Title: LGALS3BP antibody-drug conjugate enhances tumor-infiltrating lymphocytes and synergizes with immunotherapy to restrain neuroblastoma growth.

doi: 10.1186/s12967-025-06434-1

Figure Lengend Snippet: Fig. 6 Synergistic and durable anti-tumor activity of combination therapy based on anti-LGALS3BP ADC 1959-sss/DM4 and immune check-point inhibi tor anti-PD-1 in murine syngeneic neuroblastoma model. (A) Tumor growth expressed as tumor volume (mm3) from first treatment of NXS2 hLGALS3BP allografts in syngeneic A/J mice treated or not with anti-PD-1 (10 mg/kg; twice weekly; five total i.p. injections), naked 1959-sss (10 mg/kg; twice weekly; three total i.v. injections), ADC 1959-sss/DM4 (10 mg/kg; twice weekly; three total i.v. injections), or combination of 1959-sss and anti-PD-1, or combina tion of 1959-sss/DM4 and anti-PD-1. Blue and pink arrows indicate 1959-sss or 1959-sss/DM4 and anti-PD-1 treatments administration, respectively. 5 or 6 mice were considered per experimental group. (B) Kaplan-Meier curve showing probability of survival of mice bearing NXS2 hLGALS3BP allografts treated or not with anti-PD-1, 1959-sss, ADC 1959-sss/DM4, combination of 1959-sss and anti-PD-1, or combination of 1959-sss/DM4 and anti-PD-1 from first treatment. Log-rank (Mantel-Cox) test: *** p < 0.001

Article Snippet: For the evaluation of serum and EVs-associated human LGALS3BP levels in human neuroblastoma pseudometastatic experiments, human LGALS3BP ELISA Kit was performed according to manufacturer’s instructions (#KE00155; Proteintech, USA).

Techniques: Activity Assay

Journal: PLoS ONE

Article Title: Systematic Analysis of Blood Cell Transcriptome in End-Stage Chronic Respiratory Diseases

doi: 10.1371/journal.pone.0109291

Figure Lengend Snippet: A) Tree analysis of the PAH signature. Identification of the most representative genes by Gominer, PAM, IPA analysis and validation by qPCR of the candidate genes in the validation cohort; B) Network generated by IPA on the most significant GO categories in PAH signature. Over-expressed genes in PAH are in gray. C) PAM analysis based on the cluster, Green represents relatively low expression, and red indicates relatively high expression; D and E) Validation by qPCR of MDK and LGALS3 in the validation cohort, respectively.

Article Snippet: Real-time quantitative PCR was performed on a ViiA7 Fast Real-Time PCR System (Applied Biosystems) using commercially available primers: HPRT1 (Hs99999909_m1), β 2M (Hs00984230_m1), ACTB (Hs99999903_m1), TCF-7 (Hs00175273_m1), CD6 (Hs00198752_m1), IL-7R (Hs00233682_m1), LGALS3 (Hs00173587_m1), MDK (Hs00171064_m1) TLR4 (Hs00152939_m1), NLRC4 (Hs00802666_m1) and TLR8 (Hs00607866_mH).

Techniques: Biomarker Discovery, Generated, Expressing

Journal: Nature Communications

Article Title: Interactome analysis reveals that lncRNA HULC promotes aerobic glycolysis through LDHA and PKM2

doi: 10.1038/s41467-020-16966-3

Figure Lengend Snippet: a The volcano plot of the identified proteins. The proteins significantly enriched in the tobramycin purified samples are shown as red dots. Log fold change was plotted on the x -axis and −log 10 P value was plotted on the y -axis. b The protein interactome network of HULC. PPI information was obtained through a database search using String 9.0 and imported into Cytoscape 3.1.1 for network construction. Proteins and their interactions are shown as nodes and edges. Node size reflects the interaction degree. Proteins are grouped based on their known biological functions. c Validation of selected identified proteins by RIP. Immunoblots of TGM2, DDX58, LGALS3BP, H2A and PDIA3 in the cell lysates and immunoprecipitates are shown in the upper panel. The agarose gel electrophoresis images of HULC amplified by qRT-PCR are shown in the lower panel. LincRNA-p21 was used as an irrelevant lncRNA control. See also Supplementary Figs. and . Source data are provided as a Source Data file.

Article Snippet: Rabbit polyclonal antibody against PKM1 (Cat# 15821-1-AP; 1:1000), PKM2 (Cat# 15822-1-AP; 1:30 for IP), LDHA (Cat# 19987-1-AP; 1:30 for IP), LDHB (Cat# 14824-1-AP; 1:2000), TGM2 (Cat# 15100-1-AP; 1:1000), DDX58 (Cat# 20566-1-AP; 1:300), LGALS3BP (Cat# 10281-1-AP; 1:500) and mouse monoclonal antibody against FGFR1 (Cat# 60325-1-Ig; 1:1000) were from Proteintech (Chicago, IL, USA).

Techniques: Purification, Biomarker Discovery, Western Blot, Agarose Gel Electrophoresis, Amplification, Quantitative RT-PCR, Control

Journal: Microscopy research and technique

Article Title: Stromal cells of giant cell tumor of bone show primary cilia in giant cell tumor of bone.

doi: 10.1002/jemt.23976

Figure Lengend Snippet: Fig. 1. Histological diagnosis and preliminary study of GCTB. a) Hematoxylin-Eosin staining shows two cellular types in GCTB: mononuclear and multinuclear giant cells (GC). Some of the mononuclear cells showed a characteristic paranuclear vacuole (arrow). Scale bar = 50μm b) CD68 is a specific marker for monocyte-macrophage lineage. Thus, histiocytic mononuclear cells and giant cells (GC) expressed CD68. Scale bar = 50μm c) Histone H3.3 G34W mutation is a specific marker of the stromal neoplastic cells of GCTB. According to this, only mononuclear cells were stained, some of them expressing paranuclear vacuoles (arrow).Scale bar = 50μm d) Immunolabeling for Ki67, marker of cell cycle activation and proliferation, stained uniquely mononuclear cells. Scale bar = 100μm

Article Snippet: Primary antibodies used in this study were: monoclonal rabbit anti-Histone H3.3 G34W (2.5 μg/mL, Quimigen S.L., 31-1145-00, Madrid, Spain), polyclonal rabbit anti-Gli1 (1:200, ABCAM, ab49314, Cambridge, UK), polyclonal rabbit anti-Smo (1:1000, LS Bioscience, LS-B4911, Seattle, USA) and polyclonal goat anti-Patched (1:100, Santa Cruz, sc6149, Dallas, USA).

Techniques: Biomarker Discovery, Staining, Marker, Mutagenesis, Expressing, Immunolabeling, Activation Assay

Journal: Microscopy research and technique

Article Title: Stromal cells of giant cell tumor of bone show primary cilia in giant cell tumor of bone.

doi: 10.1002/jemt.23976

Figure Lengend Snippet: Fig.5. Stromal cells show primary cilia. a) Immunofluorescent co-localisation of Histone H3.3 G34W mutation in green and Arl13b (ciliary membrane) in red showed that some stromal cells present primary cilia. Scale bar = 20μm b-d) Magnification of cells showing Histone H3.3 G34W mutation and Arl13b co-localisation.

Article Snippet: Primary antibodies used in this study were: monoclonal rabbit anti-Histone H3.3 G34W (2.5 μg/mL, Quimigen S.L., 31-1145-00, Madrid, Spain), polyclonal rabbit anti-Gli1 (1:200, ABCAM, ab49314, Cambridge, UK), polyclonal rabbit anti-Smo (1:1000, LS Bioscience, LS-B4911, Seattle, USA) and polyclonal goat anti-Patched (1:100, Santa Cruz, sc6149, Dallas, USA).

Techniques: Mutagenesis, Membrane

Journal: Molecular Cancer

Article Title: An integrative model for recurrence in ovarian cancer

doi: 10.1186/1476-4598-7-8

Figure Lengend Snippet: List of gene targets selected for TaqMan ® validation in cohort 1

Article Snippet: LGALS3BP , 185853 , 9.49E-05 , 4.12 , Downregulated , Hs00174774_m1.

Techniques: Biomarker Discovery