Animals and tumor cell implantation

A) C57BL/6 (B6) and BALB/cJ mice implantation: 5 × 10⁴ E0771-luc or 4T1 cells were implanted in the mammary fat pad of seven-to eight-week-old C57BL/6 (B6) or BALB/c female mice, respectively. Ten days following implantation, mice were intraperitoneally injected for the time course of the experiment as follows: EnnA (10 mg/kg) or 17-AAG (50 mg/kg), or DMSO (control). Animal body weight and tumor volume were measured and recorded. At the end of the experiment, mice were necropsied, and tumor tissues and spleens were carefully dissected. Collected tissues were further processed for histopathological, flow cytometric, Western blotting, and RTqPCR analyses. B) Nude mouse implantation: 5 × 10⁴ E0771-luc cells were implanted in the mammary fat pad of seven-to eight-week-old female nude mice. Ten days following implantation, mice were intraperitoneally injected for the time course of the experiment with EnnA (10 mg/kg) or DMSO (control). Animal body weight and tumor volume were measured and recorded. All mouse procedures were conducted in accordance with the University Committee on the Use and Care of Animals at Augusta University.