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Transforming growth factor‐β activated kinase 1 ( TAK 1) has been shown to play a crucial role in cell death, differentiation, and inflammation. Here, we live‐imaged robust  TAK 1 activation in Lewis lung carcinoma 3 LL  cells implanted into the s.c. tissue of syngeneic C57 BL /6 mice and treated with polyinosinic:polycytidylic acid (PolyI:C). First, we developed and characterized a Förster resonance energy transfer‐based biosensor for  TAK 1 activity. The  TAK 1 biosensor, named Eevee‐ TAK 1, responded to stress‐inducing reagents such as anisomycin, tumor necrosis factor‐α, and interleukin1‐β. The anisomycin‐induced increase in Förster resonance energy transfer was abolished by the  TAK 1 inhibitor (5z)‐7‐oxozeaenol. Activity of  TAK 1 in 3 LL  cells was markedly increased by PolyI:C in the presence of macrophages. 3 LL  cells expressing Eevee‐ TAK 1 were implanted into mice and observed through imaging window by two‐photon excitation microscopy. During the growth of tumor, the 3 LL  cells at the periphery of the tumor showed higher  TAK 1 activity than the 3 LL  cells located at the center of the tumor, suggesting that cells at the periphery of the tumor mass were under stronger stress. Injection of PolyI:C, which is known to induce regression of the implanted tumors, induced marked and homogenous  TAK 1 activation within the tumor tissues. The effect of PolyI:C faded within 4 days. These observations suggest that Eevee‐ TAK 1 is a versatile tool to monitor cellular stress in cancer tissues.

cancer science

Live imaging of transforming growth factor‐β activated kinase 1 activation in Lewis lung carcinoma 3 LL  cells implanted into syngeneic mice and treated with polyinosinic:polycytidylic acid