RNA Sequencing Facilitates Quantitative Analysis of Transcriptomes of human early hematopoietic differentiation after LGR4 depletion and R-spondin2 treatment

Purpose: The goals of this study are to identify differential genes and signaling pathways after LGR4 depletion and R-spondin2 treatment through comparing thel transcriptome profilings in hESC samples collected from day2 of co-cutlure hematopoietic differentiation. Methods: mRNA profiles of hESC samples collected from day 2 of hematopoiesis differentiation were generated by deep sequencing using Illumina GAIIx. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays Conclusions: Treatment of R-spondin2 promotes up-regulation of mesoderm associated genes and LGR4 depletion severely down-reguletes mesoderm associated genes. Overall design: mRNA profiles of hESC samples collected from day 2 of hematopoiesis differentiation were generated by deep sequencing using Illumina GAIIx.