Fig. 3

EP300/CBP knockdown leads to reduced H3K27ac in APPDup and NDC neurons. A Schematic detailing RNAi knockdown during iPSC-neurons differentiation. Cells were transfected with siRNA constructs at Day 4 of neuronal induction and again on Day 8; cells and media for genetic and molecular assays were harvested on day 21. B, C siRNA knockdown of CBP (B) and EP300 (C) results in reduction of respective transcripts lasting until day 21 (p < 0.05 for all comparisons, unpaired two-tailed Ttest). Normalized read-counts were obtained by RNA-Seq. D CBP/EP300 KD result in decreased acetylation of H3K27 in both APPDup and NDC neurons at day 21. Representative Western blots of protein levels of H3K27ac and total H3. E Quantification of H3K27ac/H3 signal across three independent iPSCneuron differentiations and RNAi treatments