GSM7781675: Splenocytes_Experiment_HTO; Mus musculus; OTHER
To obtain BMDCs, femurs and tibia were removed, and BM was flushed. OT1 CD8+ T cells were isolated from the spleen of OT1 mice using EasySep™ Mouse CD8+ T Cell Isolation Kit (STEMCELL Technologies #19853). CD11c+ cells were isolated from the spleen using EasySep™ Mouse APC Positive Selection Kit II (STEMCELL Technologies #17667). B16 tumors were minced and digested using DMEM media supplemented with 2.5 mg/ml Collagenase D (Roche, #11088866001) and 0.2 mg/ml DNaseI (Roche, # 10104159001). Tumors were incubated in the water bath at 37 degrees while shaking for 10 min. After digestion samples were strained through 100-um filters and centrifuged at 1000 rpm at 4 degrees for 5 min. Cell pellets were resuspended in 1 ml of ACK lysing buffer (Gibco, #A100492-01), incubated for 5 min at RT, and washed twice with quenching buffer (PBS + 2% FCS). According to 10x genomics Chromium single-cell V3 3′ protocol. For HTO, ADT and GDO amplification we used dial-up PCR.