Gene expression analysis of E2A forced homodimer overexpression in mouse embryonic stem cells.

We set out to investigate whether overexpression of the bHLH transcription factor, E2A, is sufficient to drive neural differentiation in mouse embryonic stem cells under non-permissive culture conditions. Having identified that overexpression of specifically E2A homodimers, but not E2A monomers, was sufficient to drive neural differentiation, we sought to identify early downstream targets of E2A during the process of neural commitment using RNA-sequencing. A doxycycline-inducible cell line was used to drive overexpression of a forced homodimer construct of E2A in mouse embryonic stem cells. Cells were induced with 1ug/mL doxcycline for up to 24 hours, and samples were collected for RNA-sequencing at 0h, 18h and 24h time-points. All samples were collected in triplicate (total of 9 samples). Differential analysis was performed using the following comparisons: 0h vs. 18h; 0h vs. 24h; 18 vs. 24h.