RNA-sequencing analysis of murine Hematopoietic Stem Cells treated with Nicotinamide Riboside

LT-HSC and ST-HSCs were FACS sorted (>10^4cells per sample), cultured for two days in presence or absence of 1 mM NR and total RNA extracted Overall design: LT-HSCs (Lin-ckit+sca1+CD150+CD48-CD34-) and ST-HSCs (Lin-ckit+sca1+CD150+CD48-CD34+) were FACS-sorted and cultured for 2 days. Cultures were maintained in Stemline II (Sigma) supplemented with 100ng/ml SCF (R&D) and 2ng/ml Flt3 (R&D), +/ - 1 mM NR. NR was replenished after 24h into the culture media. At the end of the culture period, total RNAs were extracted using the Quick-RNA Microprep Kit (R1050, Zymo Reseach) following the manufacturer's instructions.