Female SCID mice or C57/BL6 micec (6-8 weeks old) were obtained from the Model Animal Research Center of Nanjing University and housed under pathogen-free conditions on a 12/12 h light/dark cycle with free access to food and water. For the MDA-MB-231 metastasis model 1.5×10 6 MDA-MB-231 cells suspended in 100μL PBS were injected into SCID mice through the tail vein.
Vehicle or 40mg/kg etomoxir was injected intraperitoneally every 2 days and mice were weighed every week before sacrificed. At the endpoint, respective metastatic organs were harvested and metastasis was confirmed by H&E staining. For the E0771 metastasis model, 5×10 5 E0771 vector or NNMToverexpression cells suspended in 100μL PBS were injected into C57/BL6 mice through the tail vein.
Cells were treated with etomoxir 2 days before injection, and vehicle or 40mg/kg etomoxir were injected intraperitoneally every 2 days. The lungs were harvested for H&E staining and IHC after 2 weeks. For the MCF-7 metatstasis assay in vivo, MCF-7 vector and NNMT-OE cells were labeled with CellTracker Green (C2925, ThermoFisher) and injected into SCID mice via the tail vein. For the etomoxir treatment group, cells were treated with etomoxir 2 days before injection. Lung tissues were collected at 4 hours and 24 hours after injection and then processed into frozen sections, which were stained with DAPI for fluorescence microscopy.
Vehicle or 40mg/kg etomoxir was injected intraperitoneally every 2 days and mice were weighed every week before sacrificed. At the endpoint, respective metastatic organs were harvested and metastasis was confirmed by H&E staining. For the E0771 metastasis model, 5×10 5 E0771 vector or NNMToverexpression cells suspended in 100μL PBS were injected into C57/BL6 mice through the tail vein.
Cells were treated with etomoxir 2 days before injection, and vehicle or 40mg/kg etomoxir were injected intraperitoneally every 2 days. The lungs were harvested for H&E staining and IHC after 2 weeks. For the MCF-7 metatstasis assay in vivo, MCF-7 vector and NNMT-OE cells were labeled with CellTracker Green (C2925, ThermoFisher) and injected into SCID mice via the tail vein. For the etomoxir treatment group, cells were treated with etomoxir 2 days before injection. Lung tissues were collected at 4 hours and 24 hours after injection and then processed into frozen sections, which were stained with DAPI for fluorescence microscopy.
