Identification of stably expressed housekeeping miRNAs in endothelial cells and macrophages in an inflammatory setting.

ABSTRACT: Reliable quantification of miRNA expression by qRT-PCR crucially depends on validated housekeepers for data normalization. Here we present thoroughly tested miRNAs eligible as references in immunological studies utilizing endothelial cells and macrophages, respectively. Endothelial cells (cell line: TIME) and macrophages (cell line: RAW264.7) were treated with various pro- and anti-inflammatory mediators (cytokines, LPS, unsaturated fatty acids) given as either single substances or in combination. Isolated RNA was screened for stably expressed miRNAs by next generation sequencing. Housekeeper candidates were thereafter validated by means of two independent quantification techniques: qRT-PCR for relative quantification and ddPCR for absolute quantification. Both methods consistently confirm

SUBMITTER: Link F 

PROVIDER: S-EPMC6726651 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature