line was maintained in C57Bl6 background, other lines were maintained in a mixed genetic background including C57BL6, FVB, and Black Swiss. Mice were housed under the following conditions: temperature: 68–74 °F; humidity: 30–60%; light cycle: 12-h light cycle at the Medical University of South Carolina and 14/10 light cycle at the Medical College of Wisconsin. Both males and females of approximately equal ratios were included in the analysis.
At the end of dissociation, normal and tumor samples were processed in the same way. 40 ml of cold DMEM + 10% fetal bovine serum was then added to the dissociation mixture. Samples were then filtered through a 70 μm membrane and then a 100μm membrane. Red blood cell lysis buffer was then added to the pellet. The pellet was resuspended and pipetted gently for 1 min. 15 ml of PBS + 10%FBS was added to the sample. After centrifuge, cells are resuspended in FACS buffer .
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