Effect of depletion of FXR on gene expression in islets from FXR knockout mice

FXR (Farnesoid X Receptor), also known as Nr1h4, is a transcription factor who is also one of the bile acid receptor has been reported to be widely involved in regulating blood glucose, bile acid, and the apoptosis of hepatocyte. We have been studing the key role of FXR in Type 2 Diabetes, which can regulate the insulin secretion or even the apoptosis of ß-cells. Thus,we measured the gene expression in the islets of FXR KO mice. Our data shows the abnormal expressed gene between the FXR KO mice and the Control group, which may be regulated by FXR. It will be significant promotion for the research in mechanism of FXR regulating the insulin secretion and apoptosis of ß-cells. Overall design: FXR gene knockout mice and corresponding control mice aged three months were selected, and the pancreatic tissue was isolated then digested by collagenase p.The total RNA of each group of islets was extracted separately.After the total RNA extracted from the sample was treated with DNase I, the eukaryotic mRNA was enriched with magnetic beads with Oligo (dT).The interrupting reagent was added to break the mRNA into short fragments, and a strand of cDNA was synthesized with six-base random hexamers using the interrupted mRNA as a template, and the second strand of cDNA was synthesized by adding buffer, dNTPs and DNA polymerase I.After kit purification and recovery, sticky end repair, adding base "A" to the 3 'end and connecting the sequencing connector, the obtained fragment was amplified and enriched by PCR after size selection.The constructed library was qualified by Agilent 2100 Bioanalyzer and ABI StepOnePlus Real-Time PCR System, and then sequenced by Illumina sequencing platform.