Transcriptomes of CD4+ Treg and non-Treg T cells of subcutaneous cell-line tumors of C57BL/6J mice with/without obesity/metformin treatment

Briefly, male, 2-3-month old, C57BL/6J mice were treated with high-fat diet for 16 weeks to induce obesity. Age/sex-matched control mice were fed normal diet. Cohorts from obese and non-obese groups (continued on same diet) were injected subcutaneously with mouse Lewis lung carcinoma cells and subsequently monitored until established tumors were noted (~day 11), when mice were injected intra-peritoneally every two days with metformin (50 mg/kg in phosphate-buffered saline [PBS]) or PBS alone. Mice were sacrificed about 3 weeks after cell injection. CD4+ Treg and non-Treg cells were collected by flow cytometry-based sorting of dissociated tumors of mice. Cells of a total of 12 tumors (12 mice) were collected. Libraries for RNA sequencing of the cells were prepared using Takara Bio SMART-Seq V4 Ultra Low Input RNA Kit. Libraries were sequenced to obtain single-end 101 b reads on a NovaSeq 6000 instrument. All work was done in one batch. There were 3 mice for each of the 4 experiment groups.