murine triple negative breast cancer tnbc cell line 4t1  (ATCC)

Journal: The Journal of Biological Chemistry

Article Title: TIM-3 inhibition enhances breast tumor progression and metastasis: A paradoxical immune checkpoint response

doi: 10.1016/j.jbc.2025.111096

Figure Lengend Snippet: Treatment with anti-TIM3 monoclonal antibody increase tumor growth and weight. A , treatment schema. Six weeks old female Balb/c mice were inoculated with 4T1 (1.2 × 10 6 ) at the fourth abdominal mammary duct of the left flank (n = 3/group). Anti-TIM3 antibody (250 μg/injection) was given i.p. on day 18,21, 24 and day 28. B and C , in vivo tumor development at day 40. B , 4T1 induced control breast tumor ( C ) Breast tumor after anti-TIM3 administration. D , representative photograph of the visceral organs of Tumor mice and anti-TIM3 treated mice collected on day 40 after tumor inoculation. E , tumor length and width were measured on day 8, 16, 21, 24, 32 and until day 40 and tumor volume were determined. Bar plot represents mean ± SD. ∗∗ p < 0.01 (Unpaired t test with Holm-Šídák method). F , tumor weight was measured on day 40, the day of resection of each group of mice. Data represent mean ± SD. ∗∗∗ p < 0.001 (Unpaired two tailed t test).

Article Snippet: Murine triple-negative breast cancer (TNBC) cell line 4T1 was purchased from ATCC (ATCC, cat. #CRL-2539, RRID: CVCL_0125) and cultured in RPMI-1640 (Himedia, cat. #AL162S) supplemented with 10% fetal bovine serum (Himedia, cat. #RM112) and 1% penicillin/streptomycin in a 37 °C incubator with 5% CO 2 and humidification.

Techniques: Injection, In Vivo, Control, Two Tailed Test

Journal: The Journal of Biological Chemistry

Article Title: TIM-3 inhibition enhances breast tumor progression and metastasis: A paradoxical immune checkpoint response

doi: 10.1016/j.jbc.2025.111096

Figure Lengend Snippet: Histopathological and immunohistochemical evaluation of TIM-3 into 4T1 allograft breast tumors and detection of liver metastases following anti-TIM-3 monoclonal antibody treatment. A and B , TIM-3 immunohistochemical detection mostly observed into tumor infiltrating immune cells (indicated by red arrow ) in primary breast tumor tissue into both control tumor mice and anti-TIM3-treated mice, respectively, at × 400 magnification with a scale bar 20 μm. C , TIM-3 expression scoring into tumor (Control) and anti-TIM-3 treated group was determined semi-quantitatively by counting the percentage of TIM-3-positive cells and staining intensity of five × 400 magnification fields selected randomly and represented in the bar graph as mean ± SD, from three mice per group, ∗∗∗ p < 0.001 (Unpaired two-tailed t-test). D-G , primary breast tumor tissue section stained with H&E, ( D and E ) breast tumor tissue histology of control tumor mice and anti-TIM3 treated mice (at × 100 magnification respectively, with scale bar 100 μm), and ( F and G ) breast tumor tissue histology of control tumor mice and anti-TIM3 treated mice respectively (at × 400 magnification respectively, with scale bar 100 μm) resected after day 40. H – L , histological analysis of liver metastasis of tumor-bearing mice and treated group showed ( H ) tumor foci (indicated with a black circle ) of control tumor mice, and ( I ) anti-TIM3 mAb-treated tumor mice showed several tumor foci (indicated with several black circles ). J , the number of tumor metastatic foci into liver tissue section was determined by counting fifty high power field under × 400 magnification of each mouse of an experimental group (n = 3). Data represent mean ± SD. ∗ p < 0.05 (Unpaired two-tailed t -test). Infiltration of inflammatory cells into liver was observed and demarcated by ( K ) blue arrow indicated the moderate (not too severe) infiltration into control tumor-bearing mice, while ( L ) the green arrow indicated the severe inflammatory cell infiltration into the liver of anti- TIM-3 treated group. All data represent mean ± SD; n = 3.

Article Snippet: Murine triple-negative breast cancer (TNBC) cell line 4T1 was purchased from ATCC (ATCC, cat. #CRL-2539, RRID: CVCL_0125) and cultured in RPMI-1640 (Himedia, cat. #AL162S) supplemented with 10% fetal bovine serum (Himedia, cat. #RM112) and 1% penicillin/streptomycin in a 37 °C incubator with 5% CO 2 and humidification.

Techniques: Immunohistochemical staining, Control, Expressing, Staining, Two Tailed Test

Journal: bioRxiv

Article Title: PEGylation strategies for enhanced nanoparticle delivery to tumor associated immune cells

doi: 10.1101/2025.07.23.666401

Figure Lengend Snippet: (A) Representative flow cytometr contour plots of NP gating strategy for live cells isolated from 4T1 tumors. Percent of (B) live cells, (C) leukocytes or tumor cells, (D) neutrophils, dendritic cells, or macrophages containing NPs obtained from tumor tissue. Percent of (E) live cells, (F) leukocytes or CD45-spleen cells, (G) neutrophils, dendriti cells, or macrophages containing NPs obtained from spleen tissue. Percent of (H) live cells, (I) leukocytes or liver cells, (J) neutrophils, dendritic cells, or macrophages containing NPs obtained from liver tissue. *p<0.05, **p<0.01, and ***p<0.001 by two-way ANOVA with Tukey correction. N=6 mice for No PEG, PEG-L5, and PEG-B10, N=4 mice for No NPs

Article Snippet: 4T1 murine triple negative breast cancer cells were obtained from ATCC (# CRL-2539, RRID:CVCL_0125).

Techniques: Isolation

Journal: bioRxiv

Article Title: PEGylation strategies for enhanced nanoparticle delivery to tumor associated immune cells

doi: 10.1101/2025.07.23.666401

Figure Lengend Snippet: (A) Representative flow cytometry contour plots of NP gating strategy for live cells isolated from 4T1 tumors. Percent of (B) live cells, (C) leukocytes or tumor cells, (D) neutrophils, dendritic cells, or macrophages containing NP obtained from tumor tissue. Percent of (E) live cells, (F) leukocytes or CD45-spleen cells, (G) neutrophils, dendritic cells, or macrophages containing NPs obtained from spleen tissue. Percent of (H) live cells, (I) leukocytes or liver cells, (J) neutrophils, dendritic cells, or macrophages containing NPs obtained from liver tissue. *p<0.05, **p<0.01, and ***p<0.001 by two-way ANOVA with Tukey correction. N=6 mice for No PEG, PEG-L5, and PEG-B10, N=4 mice for No NPs

Article Snippet: 4T1 murine triple negative breast cancer cells were obtained from ATCC (# CRL-2539, RRID:CVCL_0125).

Techniques: Flow Cytometry, Isolation

Journal: bioRxiv

Article Title: Instant fluorescence lifetime imaging microscopy reveals mechano-metabolic reprogramming of stromal cells in breast cancer peritumoral microenvironments

doi: 10.1101/2025.05.28.656717

Figure Lengend Snippet: The inclusion of 4T1 CM counteracts compression-induced metabolic reprogramming in a) NIH3T3 (data from and ) and b) 3T3-L1 cells (data from and ), but not in c) d3T3-L1 cells (data from and ). Additionally, 4T1 CM significantly increases fluorescence lifetime regardless of compression status in d) NIH3T3 (data from and ), e) 3T3-L1 (data from and ), and f) d3T3-L1 (data from and ) cells. Error bars represent SEM. Statistical significance was determined using a Student’s t-test, with asterisks indicating significance levels (* p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001).

Article Snippet: Murine 4T1 triple-negative breast cancer (TNBC) cells (ATCC, CRL-2539) were cultured in Roswell Park Memorial Institute (RPMI) medium (Corning, 10-040-CV) supplemented with 10% FBS and 1X P/S.

Techniques: Fluorescence

Journal: bioRxiv

Article Title: Instant fluorescence lifetime imaging microscopy reveals mechano-metabolic reprogramming of stromal cells in breast cancer peritumoral microenvironments

doi: 10.1101/2025.05.28.656717

Figure Lengend Snippet: Brightfield images of human TNBC with adjacent breast tissue stained a) with hematoxylin & eosin (H&E) and b) for α-SMA (fibroblast marker). c) Representative fluorescence lifetime images and associated phasor plot for mature adipocytes and fibroblasts in the tumor (left), peritumor (middle), and normal stroma (right). d) Distribution of fluorescence lifetime in tumor-associated (left), peritumoral (middle), and normal stromal (right) adipocytes and fibroblasts. Consistent with trends observed in , histogram comparisons reveal that adipocytes in the tumor exhibit lower fluorescence lifetime values, indicative of a more glycolytic phenotype, whereas those in the normal stroma display higher values, suggesting greater oxidative metabolism. Adipocyte metabolism in the peritumor, on the other hand, appears intermediate between both regions, with cells likely undergoing a glycolytic transition potentially reflecting changes in maturity states. In fibroblasts, fluorescence lifetime trends in the peritumor and normal stroma are less distinct due to tissue heterogeneity, wherein multiple stromal components in the tissue contribute to a complex, non-unimodal lifetime distribution, but generally feature glycolytic trends. Advanced deconvolution methods are therefore needed to accurately resolve metabolic activity in fibroblasts from other tissue components (e.g., matrix) within the same region. Black scale bar represents 2 mm, white scale bar is 50 µm.

Article Snippet: Murine 4T1 triple-negative breast cancer (TNBC) cells (ATCC, CRL-2539) were cultured in Roswell Park Memorial Institute (RPMI) medium (Corning, 10-040-CV) supplemented with 10% FBS and 1X P/S.

Techniques: Staining, Marker, Fluorescence, Activity Assay