GSM9392422: NTC ER-Hoxb8 macrophages grown for 7div with 0.016um baricitinib, Replicate 4; Mus musculus; RNA-Seq
For in vitro populations, RNA was extracted using the Qiagen RNeasy Mini Kit (74104) according to manufacturer's instructions. For in vivo populations, cells were isolated and FACS sorted into TRIZol LS reagent.  Cells were briefly vortexed and 0.2 volumes of chloroform (Sigma, C2432-500ML) were added. Cells were again vortexed, incubated for 3 m at RT, and then centrifuged at 12,000xg for 15 m at 4c. The upper aqueous phase was then collected and 1 volume of fresh 70% EtOH was added. Cells were again vortexed and then run through the Qiagen RNeasy Micro Kit (74004) according to manufacturer's instructions. Isolated RNA was stored at -80. Sequencing libraries were prepared in-house using either 40 ng or 8ul RNA per sample, depending on if RIN scores and concentrations were able to be calculated, using the NEBNext Single Cell/Low Input RNA Library Prep Kit for Illumina (NEB, E6420) according to manufacturer's instructions.