Gene expression profile at single nucleus level for tumor associated macrophage (TAMs) nuclei sorted from tumor of control-PyMT and S100a8-creRosa26-LSL-MYCPyMT (MYC-PyMT) mice treated with low protein diet

Nuclei of TAMs were sorted for single nucleus RNA sequencing from control-PyMT and MYC-PyMT tumor treated with low protein diet, and compare their transcriptome differences. Overall design: 7-week-old Control-PyMT and MYC-PyMT mice were lethally irradiated and reconstituted with 3 million total bone marrow cells from Fcgr1-cre-Rosa26LSL-H2B-mCherry/+ mice as described previously. Two weeks later, mice were switched to a low protein diet. 6-week post dietary switch, mice were sacrificed, and tumors were collected for immune cell isolation. TAMs were sorted based on their cell surface markers (DAPI-CD45+SiglecF-Ly6G-B220-Ly6C-F4/80+Vcam+). Nuclei were isolated with an EZ PREP kit (Sigma, Cat #NUC-101) according to the manufacturer's protocol, and intact nuclei without cell contamination were further sorted by fluorescent markers (mCherry+CD11b-SYTO9+) for downstream processes.