4t1 cells  (Jackson Laboratory)

Journal: Molecular Therapy Oncology

Article Title: Fibulin-4 expressed in metastatic breast cancer is a target of peptide-based imaging probes and experimental therapeutics

doi: 10.1016/j.omton.2026.201207

Figure Lengend Snippet: BLMP6 homes to human cancer metastases (A) Lung paraffin sections from mice without or with 4T1 metastases overlayed with AZDye555-BLMP6 (12.5 μg/mL) reveal binding specificity to metastatic cells. Endomucin IF identifies endothelial cells. Graph: data quantification, representing the average Az555 signal binding across analyzed fields ( n = 3). (B) Lung paraffin section from a mouse with MDA-MB-231 metastases overlayed with AZDye555-BLMP6 (12.5 μg/mL) reveals binding to metastatic cells. N-cadherin IF identifies AZDye555-BLMP6-bound cancer cells that underwent EMT (orange arrows) and stromal cells (green arrows). (C) In a mouse pre-grafted with GFP-expressing MDA-MB-231 metastatic cells, upon i.v. injection of AZDye555-BLMP6 (50 μg), a lung section shows a GFP-positive metastatic lesion with AZDye555-BLMP6 signal, absent in the surrounding non-tumor tissue. (D) Radiolabeled 68 Ga-BLMP6 i.v.-injected into nude mice without and with MDA-MB-231 lung metastases detected by IVIS. At 1 h, PET/CT images are scaled equally (0.4%–1.96% ID/g) to allow cross-reference throughout the study. Arrow, 68 Ga-BLMP6 in metastases; b, bladder; k, kidney. (E) Biodistribution in tissues resected from mice without and with metastases in (D), with gamma counter plotted as percentage of the injected dose per gram tissue (%ID/g tissue). For graphs, shown is mean ± SEM; ∗ p < 0.05, Student t test. Scale bars, 100 μm.

Article Snippet: Spontaneous cancer progression to metastases was modeled by injecting 2 × 10 4 4T1 cells into the mammary fat pad of 10-weeks-old female Balb/c mice (Jackson Laboratory) with a 31-gauge needle.

Techniques: Binding Assay, Paraffin Section, Expressing, Injection, Positron Emission Tomography-Computed Tomography

Journal: Molecular Therapy Oncology

Article Title: Fibulin-4 expressed in metastatic breast cancer is a target of peptide-based imaging probes and experimental therapeutics

doi: 10.1016/j.omton.2026.201207

Figure Lengend Snippet: MMAE-BLMP6 kills metastatic cells (A) Survival of 4T1 and 4T1-FBLN4-KO cells treated with indicated concentrations of MMAE-BLMP6 for 24 h measured by CellTiter-Blue cell assay. Images: trypan blue staining of 4T1 cells, untreated or treated with 1 μM MMAE-BLMP6. (B) Survival of MDA-MB-231 cells treated with indicated concentrations of MMAE-BLMP6 and MMAE-BLMP5 peptides for 24 h measured by CellTiter-Blue cell assay. Images: trypan blue staining of MDA-MB-231 cells, untreated or treated with 1 μM MMAE-BLMP6. (C) Lung paraffin section from mice with B16F10 metastases injected (sc) with 20 μg of MMAE-BLMP6 and sacrificed 24 h later. TUNEL assay identifies apoptosis in metastatic cells (arrows). (D) Lung paraffin section from mice with MDA-MB-231-luc-GFP metastases injected with MMAE-BLMP6. IF with an antibody against cleaved caspase 3 identifies apoptosis in metastatic cells (green arrows). IB4, isolectin B4 (Cy3-conjugated) binding marking the endothelium. Scale bars, 100 μm.

Article Snippet: Spontaneous cancer progression to metastases was modeled by injecting 2 × 10 4 4T1 cells into the mammary fat pad of 10-weeks-old female Balb/c mice (Jackson Laboratory) with a 31-gauge needle.

Techniques: Staining, Paraffin Section, Injection, TUNEL Assay, Binding Assay

Journal: Molecular Therapy Oncology

Article Title: Fibulin-4 expressed in metastatic breast cancer is a target of peptide-based imaging probes and experimental therapeutics

doi: 10.1016/j.omton.2026.201207

Figure Lengend Snippet: MMAE-BLMP6 suppresses metastasis progression (A) Kaplan-Meier curves showing cumulative survival of C57BL/6 mice i.v.-grafted with B16F10-luc cells and subcutaneously (sc)-injected with MMAE-BLMP6 or PBS as indicated. (B) Bioluminescence analysis of mice from (A) by IVIS revealing metastasis suppression in MMAE-BLMP6-treated mice (days 15 vs. 5). (C) Kaplan-Meier curves showing cumulative survival of Nude-Foxn1nu mice i.v.-grafted with MDA-MB-231-luc-GFP cells and sc-injected with MMAE-BLMP6 or PBS as indicated. (D) Bioluminescence analysis of mice from (D) by IVIS revealing metastasis suppression in MMAE-BLMP6-treated mice (days 21 vs. 12). (E) Mammary tumor growth in BALB/c mice bilaterally grafted with 4T1-luc cells and sc-injected with MMAE-BLMP6 ( N = 8) or scrambled MMAE-BLMP6 ( N = 7) as indicated in (G). Plotted are mean ± SEM; ∗ p < 0.01, Student t test. (F) Bioluminescence analysis of representative mice from (E) by IVIS at week 3 reveals smaller primary tumors (PT) and metastases (M) in mice treated with MMAE-BLMP6. (G) Kaplan-Meier curves showing survival of BALB/c mice orthotopically grafted with 4T1-luc cells and sc-injected with MMAE-BLMP6 or scrambled MMAE-BLMP6 as indicated. (H) Bioluminescence analysis of mice from (F and G) by IVIS revealing metastasis growth suppression in MMAE-BLMP6-treated mice (days 33 vs. 26). (I) Representative lungs from mice that died on the same day showing smaller metastases (arrowheads) in animals treated with MMAE-BLMP6.

Article Snippet: Spontaneous cancer progression to metastases was modeled by injecting 2 × 10 4 4T1 cells into the mammary fat pad of 10-weeks-old female Balb/c mice (Jackson Laboratory) with a 31-gauge needle.

Techniques: Injection

Journal: Molecular Therapy Oncology

Article Title: Fibulin-4 expressed in metastatic breast cancer is a target of peptide-based imaging probes and experimental therapeutics

doi: 10.1016/j.omton.2026.201207

Figure Lengend Snippet: BLMP6 mimics LTBP4 and binds to FBLN4 (A) AlphaFold3 (AF3) modeling of the 3D structure of BLMP6 and mouse LTBP4L (residues 716–733 shown) containing the BLMP6 similarity segment (725–730, highlighted). (B) Left, consensus docking pose predicted by AF3 and CB-Dock shows that BLMP6 interacts with the C terminus of mouse FBLN4 (residues 41–443 shown). Right, a similar binding predicted for a 106 amino acid fragment of LTBP4L (residues 675–780) containing the BLMP6 similarity segment (725–730, highlighted) using AF3 and ClusPro. (C) Biolayer interferometry assay performed with Octet to measure binding affinities. (D) Affinity purification of FBLN4 from 4T1 cell extract with biotinylated BLMP6 immobilized on streptavidin-coated beads. Immunoblotting with anti-FBLN4 antibodies identifies the expected 49 kDa protein, which is not isolated on control beads. (E) Graph: real-time PCR analysis of gene messenger RNA (mRNA) expression, relative to 18S , confirming the knockout of FBLN4 in 4T1 cells. Images: lung paraffin sections from mice that were injected with phage displaying BLMP6. Anti-phage IF demonstrates BLMP6-phage homing to 4T1 metastases but not to 4T1-FBLN4-KO metastases. Scale bars, 100 μm.

Article Snippet: Spontaneous cancer progression to metastases was modeled by injecting 2 × 10 4 4T1 cells into the mammary fat pad of 10-weeks-old female Balb/c mice (Jackson Laboratory) with a 31-gauge needle.

Techniques: Binding Assay, Affinity Purification, Western Blot, Isolation, Control, Real-time Polymerase Chain Reaction, Expressing, Knock-Out, Injection

Journal: Communications Biology

Article Title: Aging promotes a RAGE-dependent increase in breast cancer metastasis

doi: 10.1038/s42003-026-10022-4

Figure Lengend Snippet: Transwell invasion assays using Py8119 ( A ) and 4T1 ( B ) breast cancer cells were performed using 1% serum pooled from three mice per group (young RAGE⁺/⁺, aged RAGE⁺/⁺, or aged RAGE −/− ) as the chemoattractant, and invasion was quantified after 24 h. Transwell assays assessing migration ( C ) and invasion ( D ) of Py8119 cells toward 1% serum pooled from aged RAGE⁺/⁺ mice ( n = 3) were performed in the presence of the RAGE inhibitor TTP488 (1 μM), the S100A8/9 inhibitor paquinimod (25 μM), the CCR1 inhibitor BX471 (10 μM), or the CCR2 inhibitor BMS-CCR2-22 (10 μM). Data represent mean ± SEM from four independent biological experiments. Statistical analysis: one-way ANOVA with Dunnett’s multiple-comparisons test. * p < 0.05; ** p < 0.01; *** p < 0.001; and **** p < 0.0001.

Article Snippet: 4T1 cells were purchased from ATCC and cultured in RPMI 1640 supplemented with 10% FBS (cat# 900-108, Gemini).

Techniques: Migration

Journal: Bone Research

Article Title: The endosteal niche regulates breast cancer cell dormancy in bone: identification of new molecular determinants

doi: 10.1038/s41413-026-00535-3

Figure Lengend Snippet: Expression of Notch1 and Notch2 in breast cancer cells and interaction with SNOs. a Human MDA-MB231, b human ZR75D, c mouse 4T1, d human T47D and e human BT747 BrCa cell lines were analysed by flow cytometry for their expression of Notch1 and Notch2. f Quantification of human MDA-MB231 GFP + BrCa cells (MDA GFP ) MACS-sorted for low or high expression of Notch1 after 72 h of culture on SNO and NON-SNO monolayers. g Quantification of human MDA-MB231 GFP + BrCa cells (MDA GFP ) MACS-sorted for low or high expression of Notch2 after 72 h of culture on SNO and NON-SNO monolayers. h Quantification of human T47D BrCa cells labelled with the PKH26 membrane dye, MACS-sorted for low or high expression of Notch1 after 72 h of culture on SNO and NON-SNO monolayers. Results are ( a – e ) representative or ( f – h ) the mean ± SD of 3 independent experiments. Statistics: unpaired t -test

Article Snippet: The human BrCa cell lines MDA-MB231, T47D, ZR75D, BT474, and the murine BrCa cell line 4T1 were purchased from the American Type Culture Collection (ATCC) and cultured according to the supplier instruction.

Techniques: Expressing, Flow Cytometry, Membrane