GSM8146610: BMDM, control, scRNAseq; Mus musculus; RNA-Seq
BMDMs that had successfully undergone 7 days of differentiation in 20 ng/mL M-CSF were used. Spent media was removed, BMDMs were gently washed once with phosphate-buffered saline (PBS), and fresh IMDM supplemented 10% FBS, 1% P/S, and 20 ng/mL M-CSF was added. Then, conditioned media from B16F10 cells treated with either MPS1i or DMSO was collected. Conditioned media was centrifuged for 5 min at 300 x g to remove any cellular debris. The supernatant was collected and then supplemented to 5% FBS, 1% P/S, and 20 ng/mL M-CSF. One whole volume of conditioned media was added to BMDMs with fresh media. 24 h after treatment, BMDMs were detached using 0.05% Trypsin and processed for single-cell RNA-sequencing. RNA libraries were prepared using the 10X Genomics Chromium Single Cell Gene Expression kit (v3.1, single index, PN-1000128; PN-1000127; PN-1000213) per the manufacturer's instructions.