RNA-Seq of FACS sorted Enteric Glia cells from the LMMP after T-cell activation via CD3
RNA-Seq of FACS sorted Enteric Glia cells from the LMMP after T-cell activation via CD3
The mice received a peritoneal injection of an anti-CD3 antibody or its isotype control antibody. At 6 hours, the small intestine was collected and the longitudinal muscle layer with the myenteric plexus stripped off. After enzymatic dissociation and trituration, the enteric glia were FACS sorted and finally the RNA isolated.  Total RNA was extracted from the samples taken from colons of mice using the peqGOLD total RNA kit (Peqlab/VWR) following the manufacturers' instructions. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using either dUTP for directional library or dTTP for non-directional library.The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries will be pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.