Microglial BV2-derived exosomal microRNA sequencing in heatstroke. Microglial BV2-derived exosomal microRNA sequencing in heatstroke

We performed exosomal microRNA sequencing on BV2 in heatstroke, to shed light on the expression and function changes of microglia in heatstroke. We then performed gene expression profiling analysis using data obtained from microRNA-seq of 2 groups. Overall design: Culture dishes containing BV2 cells in 37℃ cell incubator for 2h as a control group, and the experimental group are maintained in 42℃ for 2h, and then rewarmed in 37℃ for 12h.Extraction of BV2-derived exosomes by ultracentrifugation. Exosomal RNA Extraction TRIzol reagent is used to extract the RNA from exosomes. Illumina HiSeqTM 2500 is applied for sequencing, with a data volume of 20 M reads per sample.The total RNA or purified sRNA fragment of the sample is extracted. The 3'-end and 5'-end junctions are ligated, reverse transcribed into cDNA, and then amplified by PCR.During the QC step, Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System are used to qualify and quantify of the sample library.