Ten male BALB/c nude mice (19–22 g, 6 weeks old) were obtained from the Animal Center of Chinese Academy of Science (Shanghai, China). They were randomly divided into two groups of five each and housed three per cage in pathogen-free conditions at 28 °C, 50% humidity and were housed in a specific sterile environment suitable and regularly observed. The experimental protocol was approved by the Committee on the Ethics of Animal Experiments of Hainan General Hospital. SKBR-3-TR cells (1 × 107) that were stably transduced with sh-TINCR or sh-NC were subcutaneously injected into the flanks. The mice were housed for 25 days, then the formed tumors were stripped and the tumor mass was measured.
Experimental lung metastases were induced by injections of single-cell suspension (2 × 106 cells in 100 μl) into the mouse lateral tail vein. Cells were stably transduced with sh-TINCR or sh-NC, and all cell injections were administered in a total volume of 500 μL PBS containing 0.1% BSA over a 60 s duration [26 (link)]. Five weeks later, prior to in vivo imaging, the mice were anaesthetized with phenobarbital sodium and the established lung metastases images were observed by LB983 NIGHTOWL II system (Berthold Technologies GmbH, Calmbacher, Germany).
Experimental lung metastases were induced by injections of single-cell suspension (2 × 106 cells in 100 μl) into the mouse lateral tail vein. Cells were stably transduced with sh-TINCR or sh-NC, and all cell injections were administered in a total volume of 500 μL PBS containing 0.1% BSA over a 60 s duration [26 (link)]. Five weeks later, prior to in vivo imaging, the mice were anaesthetized with phenobarbital sodium and the established lung metastases images were observed by LB983 NIGHTOWL II system (Berthold Technologies GmbH, Calmbacher, Germany).