4t1 breast cancer cell lines  (ATCC)

Journal: bioRxiv

Article Title: Targeting CD73-A 2a R-Mediated Adenosine Signaling at the Tumor-Immune Interface Overcomes Radioresistance

doi: 10.64898/2026.05.26.727904

Figure Lengend Snippet: Intra-tumoral adenosine concentration (Mean ± SEM) in (A) human and (B) murine breast tumors and normal breast tissue quantified by LC-MS. (C) IHC staining: Magnification 200 X, Scale 100 μM and (D, E) H-score (Mean ± SEM) of CD73 and A 2a R in human normal breast tissue (epithelial cells, immune cells) and breast tumors (cancer cells, immune cells). (F) Relative expression of genes in the adenosine signaling axis in EO771 and 4T1 murine breast cancer cells compared to GAPDH by qPCR. (G) Frequency of immune cell subtypes (Mean ± SEM) in EO771 tumors by spectral flow cytometry. (H) Frequency of immune cell subtypes (Mean ± SEM) in 4T1 tumors by spectral flow cytometry. (I) Abundance of adenosine signaling proteins (Mean ± SEM). in CD45 - compartment of EO771 tumors. (J) Abundance of adenosine signaling proteins (Mean ± SEM). in CD45 - compartment of 4T1 tumors. (K) Expression of adenosine signaling proteins on CD45 + immune cells (Mean ± SEM) in EO771 tumors quantified using spectral flow cytometry. (L) Expression of adenosine signaling proteins on CD45 + immune cells (Mean ± SEM) in 4T1 tumors quantified using spectral flow cytometry. Tumors were harvested from n=10-12 mice, 25 days post-implantation. (myeloid cells (CD45 + CD11b + TCRβ - ), T cells (CD45 + CD11b - TCRβ + ), B-cells (CD45 + CD11b - CD19 + ), dendritic cells (CD45 + CD11b - CD11c + CD103 + ), NK cell – natural killer cells (CD45 + CD11b - CD11c - NK1.1 + ).

Article Snippet: Murine luminal B breast cancer cell line EO771 (ERα-, ERβ+, PR+, HER2+) and triple negative breast cancer cell line 4T1 (ER-, PR-, HER2-) was procured from ATCC.

Techniques: Concentration Assay, Liquid Chromatography with Mass Spectroscopy, Immunohistochemistry, Expressing, Flow Cytometry

Journal: bioRxiv

Article Title: Targeting CD73-A 2a R-Mediated Adenosine Signaling at the Tumor-Immune Interface Overcomes Radioresistance

doi: 10.64898/2026.05.26.727904

Figure Lengend Snippet: (A) Schematic summary of treatment schedule for EO771 or 4T1 orthotopic breast cancer model. (B) Tumor volume measurements (Mean ± SEM) beginning 8 days after EO771 implantation (n=8-10 mice/group). AB680 (quemliclustat, CD73 inhibitor), AB928 (etrumadenant, A 2a R/A 2b R antagonist), αPD1 (zimberelimab, anti-PD-1 antibody), isotype control (IgG2 antibody), tumor irradiation (RT, 8 Gy x 3; 12-, 13-, 14- days after implantation). (C) Tumor volumes from days 8-42 of irradiated mice treated in combination with isotype control, anti-PD-1, AB928 and AB680, AB928 and AB680 and anti-PD-1. (D) Spider plots of tumors from mice belonging to all treatment groups implanted with EO771 cells. (E) Slopes (Mean ± SEM) during tumor rebound period (days 32-42) from growth curves of mice belonging to irradiation and anti-PD-1, irradiation and AB680 and AB928, irradiation and AB680 and AB928 and anti-PD-1 treatment groups. (F) Tumor volume measurements (Mean ± SEM) beginning 8 days after 4T1 implantation. Data are representative of two separate experiments, n=8-10 mice/group (*) p<0.05 (**), p<0.01, (****) p<0.0005.

Article Snippet: Murine luminal B breast cancer cell line EO771 (ERα-, ERβ+, PR+, HER2+) and triple negative breast cancer cell line 4T1 (ER-, PR-, HER2-) was procured from ATCC.

Techniques: Control, Irradiation

Journal: Cancers

Article Title: Therapeutically Engineering Exosomes to Target CD206+ M2 Macrophages to Prevent the Development of Primary Tumors and Distal Metastases in Breast Cancers

doi: 10.3390/cancers18101619

Figure Lengend Snippet: Differential biodistribution of exosomes and protein in 4T1 tumors (Balb/c mice): I-131-labeled control exosomes (HEK293 exo), anti-mouse CD206 antibody (CD206 Ab), fusion protein (fusion P) containing targeting peptide and Fc-mIgG2b, and engineered exosomes (Engi. Exo) carrying targeting peptide were administered IV into 4T1 breast cancer-bearing animals; SPECT images were obtained 3 h after administration. ( A ) SPECT images showed the distribution of the agents in the primary tumors (white circles) and lungs (yellow rectangular areas). White arrows show possible metastatic foci. Fusion protein and engineered exosomes accumulated in the primary and metastatic tumors. We used rainbow color to depict the activity of I-131. Blue is the lowest and red is the highest activity of I-131. ( B ) Randomly selected lung H&E staining showed multiple metastatic foci (black arrows). ( C ) Presence of numerous CD206+ cells is seen in tumors (green cells). n = 2–3 animals per group. Ab = antibody, Engi = engineered, Exo = exosomes, HE = Hematoxylin and Eosin, P = protein, SPECT = single-photon emission computed tomography, H = heart.

Article Snippet: Syngeneic, aggressive triple-negative breast cancer (TNBC) mouse cell lines 4T1 (for Balb/c mice, ATCC, Manassas, VA, USA) and AT3 (for C57BL/6, Millipore, Burlington, MA, USA) were obtained and maintained in the laboratory in Dulbecco’s Modified Eagle Medium (DMEM) (Corning, Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS) (Nalgene-GIBCO, Rochester, NY, USA), sodium pyruvate (GIBCO, Grand Island, NY, USA), non-essential amino acids (GIBCO), sodium glutamate (GIBCO), and 100 U/mL penicillin and streptomycin (GIBCO).

Techniques: Labeling, Control, Single Photon Emission Computed Tomography, Activity Assay, Staining

Journal: Cancers

Article Title: Therapeutically Engineering Exosomes to Target CD206+ M2 Macrophages to Prevent the Development of Primary Tumors and Distal Metastases in Breast Cancers

doi: 10.3390/cancers18101619

Figure Lengend Snippet: Resected model of breast cancer (4T1 in Balb/c mice) and metastasis: Representative images from tumor-resected animals (two from each group). All the primary tumors were resected 11 days after tumor cell implantation, and the animals were followed up to 63 days after resection. The figure shows representative animals from the control (HEK293 ( left panel )) and engineered exosome-treated ( right panel ) groups. The animals treated with the control exosomes began showing local recurrence and lung metastasis (black arrows show local recurrence and multiple distal metastases) starting on day 28 post-resection. None of the animals that received the engineered exosomes to deplete CD206+ cells showed any recurrence or distal metastasis. It is noteworthy that a small leftover tumor (white arrow) in one of the engineered exosome-treated animals disappeared following therapy on subsequent bioluminescent imaging. n = 5 per group. Pre-Sur = pre surgery, POD = post-operative day.

Article Snippet: Syngeneic, aggressive triple-negative breast cancer (TNBC) mouse cell lines 4T1 (for Balb/c mice, ATCC, Manassas, VA, USA) and AT3 (for C57BL/6, Millipore, Burlington, MA, USA) were obtained and maintained in the laboratory in Dulbecco’s Modified Eagle Medium (DMEM) (Corning, Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS) (Nalgene-GIBCO, Rochester, NY, USA), sodium pyruvate (GIBCO, Grand Island, NY, USA), non-essential amino acids (GIBCO), sodium glutamate (GIBCO), and 100 U/mL penicillin and streptomycin (GIBCO).

Techniques: Control, Imaging