p53 is activated in ES cells after IR. R1 stem cells passaged once (P1) or twice (P2) without feeders and 3T3 mouse fibroblasts were irradiated with 7.5 Gray or left unirradiated. Cells were harvested at the indicated times and divided into two aliquots. One of the aliquots was used to determine the amount of protein (A) and the second aliquot (B) to monitor the abundance of corresponding RNAs. (A) Western Blots showing p53, p21, Bax, Noxa, Mdm2, Puma, Oct4, Nanog and PCNA (proliferating cell nuclear antigen) for loading control. (B) Real time PCR for mdm2, p21, puma, noxa, bax, oct4 and nanog RNA. Mean values and standard deviations were calculated from the obtained 2^dCT numbers of qRT-PCR signals of 3–6 independent experiments and blotted. Values of non-irradiated P2 cells at time point "0" and non-irradiated 3T3 cells, respectively, were set to 1. The quality of the qRT-PCR products was controlled by agarose gel electrophoresis. A representative picture of the gel is placed below the graph.
Solozobova et al. BMC Cell Biology 2009 10:46 doi:10.1186/1471-2121-10-46