Transcription factor Hoxb5 reprograms B cells into functional T lymphocytes [RNA-seq2]

Here we were performed RNAseq to analyze the transcriptome changing of Hoxb5 in Pro/pre-B by over expression. B220+ enriched cells from bone marrow were used to transduce Hoxb5 virus or GFP control virus. And 1000 transduced Pro/pre-B cells were sorted for RANseq analysis. The cDNA of sorted 1000-cell aliquots were generated and amplified as described previously (PMID；20203668). Then quality controls of the amplified cDNA were examined by qPCR analysis of housekeeping genes (B2m, Actb, Gapdh, Ecf1a1). RNAseq libraries of 1000 cells were generated by Nextera XT DNA Sample Preparation Kit (FC-131-1096) with 1 ng of generated dsDNA following the manufacturer’s protocol. The samples passed quality control were used for sequencing library preparation by illumine Nextera XT DNA Sample Preparation Kit (FC-131-1096). All libraries were sequenced by illumina sequencers NextSeq 500 (illumina).