RNAseq for the identification of genes induced upon TGR5 stimulation in intestinal stem cells

Purpose: Transcriptomic exploration for the identification of genes induced upon TGR5 stimulation in intestinal stem cells Methods: For each biological replicate, GFPhi cells were isolated by FACS from intestines from 4 pooled Lgr5-eGFP-IRES-CreERT2 mice. About 200.000 GFPhi cells were then embedded in Matrigel (20.000 per well in 10µL Matrigel drop) and after 4 hours were treated with INT-777 (30µM) or DMSO as control. After 16 hours RNA was extracted (RNAqueous Kit, Thermofisher) for RNAseq analysis. This experiment was repeated 3 times in 3 different days (n=3 biological replicates). Results: INT-777 treatment results in the activation of a panel of genes showing high enrichment of TEADs family of TF in associated motifs. Design: For each biological replicate, GFPhi cells were isolated by FACS from intestines from 4 pooled Lgr5-eGFP-IRES-CreERT2 mice. About 200.000 GFPhi cells were then embedded in Matrigel (20.000 per well in 10µL Matrigel drop) and after 4 hours were treated with INT-777 (30µM) or DMSO as control. After 16 hours RNA was extracted (RNAqueous Kit, Thermofisher) for RNAseq analysis. This experiment was repeated 3 times in 3 different days (n=3 biological replicates).