Fig. 3

siRNA targeting of NuRD subunits reactivates ectopic L1. HeLa cells were transfected with a synthetic human L1 retroelement cloned into a pGL4.13 backbone vector where L1 ORF1p was tagged with EGFP fluorescent protein as a marker of reactivation. Following transfection, cells were grown for 7 days until no fluorescence was detected due to epigenetic silencing. Cells were then transfected with a mixture of siRNAs targeting Mi2-β, RbAp46 and HDAC1 compared to a non-target scrambled sequence. Images were acquired 48 h post transfection. Each experiment was performed in triplicate and images shown representative of the respective fields