TGFB1 and adenocarcinoma: To evaluate the effects of mutations at Vps13d codon 295 and Tgfbr2 codon 549, quantitative real-time PCR for Vps13d and Tgfbr2, and immunoblot analysis for SMAD2/3, which was a transcription factor in TGFβ-SMAD signalling pathway, were conducted using the DMBA-treated organoid-derived adenocarcinomas, with reference to normal mammary tissues and mammary adenocarcinomas obtained in an in vivo DMBA-treated experiment using BALB/c-Trp53 knockout female mice (Machida and Imai, 2021).