Protocol for KAT8 conditional knockout mice generation and its application in 4NQO-induced esophageal tumor model

4-Nitroquinoline 1-oxide (4NQO), a quinoline derivative, induces tumor formation in mice following a multi-stage pattern analogous to human tumors, mimicking the natural carcinogenesis process. Here, we present a protocol for inducing esophageal squamous cell carcinoma (ESCC) in mice using 4NQO. We detail the acquisition of KAT8 esophagus-specific knockout mice, the establishment of 4NQO-induced ESCC, and the validation of the model via western blotting (WB), hematoxylin and eosin (H&E) staining, and immunohistochemistry (IHC). For complete details on the use and execution of this protocol, please refer to Zhang et al. 1 • Step-by-step guide for breeding of esophageal-specific KAT8 knockout mice • Protocol for 4NQO-induced esophagus tumorigenesis in KAT8-specific knockout mice • Details to verify model construction via H&E staining and IHC techniques Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 4-Nitroquinoline 1-oxide (4NQO), a quinoline derivative, induces tumor formation in mice following a multi-stage pattern analogous to human tumors, mimicking the natural carcinogenesis process. Here, we present a protocol for inducing esophageal squamous cell carcinoma (ESCC) in mice using 4NQO. We detail the acquisition of KAT8 esophagus-specific knockout mice, the establishment of 4NQO-induced ESCC, and the validation of the model via western blotting (WB), hematoxylin and eosin (H&E) staining, and immunohistochemistry (IHC).