Figure 3

shCITED2-mediated cell death is triggered by a p53-dependent pathway. (a) Gene set enrichment analysis (GSEA) of genome-wide gene expression from CD34⁺ cord blood cells transduced with a control- or shCITED2 vector revealed an enrichment of p53 target genes. (b) Q-PCR of p53 targets in NB4 and MOLM-13 cells 4–5 days after transduction with the shCITED2 vector. Error bars indicate s.d. of triplicates from representative Q-PCR. (c) MOLM-13 cells were double-transduced with GFP and mCherry expressing shRNA vectors to knockdown CITED2 and/or p53 and stained for Annexin V 4–5 days after transduction. Representative FACS plots indicating the percentage of Annexin V-positive cells are shown. (d) Percentage of Annexin V-positive cells in shCITED2/shp53-transduced MOLM-13 (n=5) and K562 (n=3) cells 5 days after transduction is shown. Error bars indicate s.d. *P<0.05, ***P<0.001; n.s.= not significant. (e) MOLM-13 cells were co-transduced with a construct for CITED2 knockdown and p53 overexpression. Percentage of Annexin V-positive cells 3 days after transduction is indicated