GSM7676665: Renca VHL KO.1 PMN MDSC LY6G M2; Mus musculus; RNA-Seq
For single cell experiments, 10-week-old Balb/c mice were injected s.c. with 1 × 106 Vhl WT or Vhl KO cells. Tumors were harvested at 16 days post injection and then enzymatically digested. CD45+ and CD45- live cells were flow sorted and counted using trypan blue and a TC20 counter. For single nucleus RNA seq experiment, nuclei was isolated from human patient ccRCC tumors as previously described [Wu., H. et al., J Am Soc Nephrol, 2019]. For bulk RNA-seq, CD11b+ fractions from experimentally matched tumors were bead isolated prior to staining of indicated surface markers and viability dye. The fractions were then sorted on a BD FACSAria III cell sorter and stored in 5 parts RNA cell protect to 1 part FACS solution. RNA was isolated, and low-input RNA sequencing was performed using the SMART-Seq v4 Ultra Low Input RNA Kit. Libraries were made according to manufacturers's protocol (10X genomics or Truseq stranded library kit) scRNAseq, snRNAseq, RNAseq