Instrument: Ion Torrent Proton
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: This osteoclast culture was performed on collagen-coated dishes, which permitted a harvesting of the adherent cells by enzymatic treatment and then a preparation of the single-cell suspensions. Murine bone marrow cells were cultured with M-CSF for 2 days, and these cells were further cultured for 3 days with RANKL in the presence of M-CSF. scRNA-seq, bulk RNA-seq and proteome analyses were performed at the following time points: Day 0 (before RANKL stimulation), Day 1 (1 day after RANKL stimulation) and Day 3 (3 days after RANKL stimulation). Bulk-RNA seq analysis was also performed using control and Cited2-KO cells on Day 3 of the osteoclastogenic culture system to investigate the role and point of action of Cited2 in the trajectory of osteoclastogenesis. RNA libraries were prepared for sequencing using standard Ion Torrent Proton protocols