GSM5570832: KO; Mus musculus; RNA-Seq
We sorted CD45+ immune cells from Kcnj2f/f and Kcnj2CKO mice and performed 10X single cell sequencing to illustrate the impact of Kcnj2 on TME. CD45+ tumor-infiltrating immune cells were enriched by FACS from dissociated day 10 B16F10 tumors for single-cell analysis. Tumors from three different mice were pooled per sample. The overall cell viability was confirmed by trypan blue exclusion with Countess II Automated Cell Counter and concentration was adjusted to 700-1200 cells/uL. Single-cell suspensions were loaded to 10x Chromium to capture ~6000-8000 single cell according to the manufacturer's instructions of 10X Genomics Chromium Single-Cell 3' kit (V3). The following cDNA amplification and library construction steps were performed according to the standard protocol. Libraries were sequenced on an Illumina NovaSeq 6000 sequencing system (paired-end multiplexing run,150bp) by LC-Bio Technology co.ltd., (Hangzhou,China) with average depth of ~48000 reads per cell.