Effect of rapamycin on murine subcutaneous MC38 tumor cell populations

To determine differential metabolic gene expression in distinct cell populations from subcutaneous MC38 tumors at baseline and in response to rapamycin, tumors from mice treated with vehicle or rapamycin were harvested, reconstituted to single cell suspensions, and flow sorted into cancer cell (CD45-), tumor-associated macrophage (TAM, CD45+ CD11b+ Ly6G- Ly6C lo F4/80 hi), monocytic myeloid-derived suppressor cell (M-MDSC, CD45+ CD11b+ Ly6G- Ly6C hi), CD8 T cell (CD45+ CD3+ CD8a+), and CD4 T cell (CD45+ CD3+ CD8a-) populations. RNA was extracted and transcripts were quantified by the NanoString nCounter Metabolic Pathways Panel. There are 3 biological replicates for all cell populations including whole tumor single cell suspension (6 cell populations) and vehicle (control) and rapamycin treatment (2 treatments) except for M-MDSC +rapamycin and CD4 +rapamycin, which have 2 biological replicates due to QC concerns in discarded samples. Therefore, in total there are 6 cell populations x 2 treatments x 3 replicates -2 samples = 34 samples