miRNomes of haematopoietic stem cells and dendritic cells identify miR-30b as a regulator of Notch1

Su, Xiaoping; Qian, Cheng; Zhang, Qian; Hou, Jin; Gu, Yan; Han, Yanmei; Chen, Yongjian; Jiang, Minghong; Cao, Xuetao

In summary, through Illumina sequencing, we present a more comprehensive understanding of the miRNomes during DC development and differentiation. miRNA expression profiles change significantly, especially at the developmental stage from HSCs to imDCs as well as the differentiational stage from maDCs to DCreg, while they are relative stable during the DC maturation process. Many stage-specific miRNAs undergo epigenetic reprogramming in their promoter regions, suggesting their specific functions in specific stages. Among them, miR-30b is an enriched miRNA in DCreg, which contributes to the preferential IL-10 and NO production in diffDCs through targeting Notch1. Our results provide a resource for researching the roles of miRNAs in DC development, differentiation and function. In addition, miRNome of mouse bone marrow HSCs may also provide clues for the future study of stem cell biology. Recombinant mouse GM-CSF, IL-4, LPS (Escherichia coli, O26:B6) and Delta 4 were from Sigma (St Louis, MO, USA). All primers were supplied by Sangon (Shanghai, China). Antibody against Notch1 (D1E11) was from Cell Signaling Technology (Danvers, MA, USA), antibodies against b-actin and their respective horseradish peroxidase-coupled secondary antibodies were from Santa Cruz (Santa Cruz, CA, USA), antibodies against Lineage(Lin), c-Kit and Scal were from eBioscience (San Diego, CA, USA). g-secretase inhibitor X (g-SiX) was from Calbiochem (San Diego, CA, USA).