RNA sequencing analyses of fused heterokaryons between mouse embryonic stem cells and human B lymphocytes collected at early timepoints after fusion

Tcf3 is an effector of Wnt canonical pathway and deletion of Tcf3 in mouse embryonic stem cells (mESCs) strongly enhances reprogramming efficiency after fusion. We fused murine Tcf3-/- mESCs with human B-lymphocytes using the Polyethylene glycol (PEG) reagent and isolated heterokaryons at different time points to analyze early reprogramming events starting from 4 hours after fusion. We performed a genome-wide expression profile of the heterokaryons by RNA-sequencing at a high resolution. Tc3-/- mESCs and human B lymphocytes labeled with distinct dyes were fused and hybrids were sorted at 4h, 12h, 48h and 120h after fusion. Human B lymphocytes mock treated with PEG were used as reference. The experiments were done in triplicates for each timepoint