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