Fig. 1.

TBX3iso1 possess enhanced tumorigenic potential in nude mice. a Total TBX3, TBX3iso1, and TBX3iso2 mRNA expression was assessed by qRT-PCR, normalized to GAPDH expression levels, and depicted as fold change relative to the empty vector control. b Cells (1.0 × 10⁷) were injected into the mammary fat pad of nude mice. Mice were sacrificed when the tumor volume reached 2500 mm³, or 1-year post injection, whichever occurred first. The Kaplan–Meier plot shows disease-free survival over 365 days post injection. An event was defined as tumor volume reaching the 2500 mm³ end-point. c Tumor volume showing growth kinetics over time. TBX3iso1 tumors are shown in various colors, while the TBX3iso2 tumor is shown in black. d Histological analysis of H&E stained slides was conducted by an anatomical pathologist for all mammary fat pads and primary tumors. Cases with atypical ductal hyperplasia (ADH), ductal carcinoma in situ (DCIS), invasive mammary carcinoma (IMC), and metastasis were documented. All tissues and lymph nodes collected were examined for metastases in mice with IMC. Metastases were confirmed through positive immunohistochemical staining for anti-human mitochondria. e Representative images of cases of DCIS, IMC showing invasion into skeletal muscle, and metastasis into the right brachial lymph node. Suspect metastases from H&E slides were confirmed through immunohistochemical staining with anti-human mitochondrial antibody. *p < 0.05, **p < 0.01, ***p < 0.001 by one-way ANOVA with Tukey post-hoc for comparison between three groups. Survival analysis for Kaplan–Meier curve was calculated using the log-rank test statistic. Error bars represent standard deviation