Increased p53 protein expression in irradiated <i>Egr1</i>^<i>+/+</i> and <i>Egr1</i>^-/- BM-MNCs is independent of mRNA expression.

<p><b>(A)</b> Cell lysates from irradiated Lin–ve enriched BM-MNCs (2 Gy or 6 Gy) at different time points (15’, 30’ and 45’) after radiation exposure were used for immunoblotting for p53. A549 and H358 cell lysates were used as positive and negative controls respectively. (<b>B)</b> Intensities of p53 bands were normalized to β-actin. Values are expressed as densitometric ratios. (<b>C)</b> BM-MNCs were lysed 45’ or 60’ after irradiation (2 Gy or 6 Gy). The RNA extracted from these lysed cells was used to determine p53 mRNA expression by qRT-PCR. (<b>D</b>) Cell lysates from phorbol 12-myristate 13-acetate (PMA) stimulated (30ng/ml) or irradiated Lin–ve encriched <i>Egr1</i>^<i>+/+</i> BM-MNCs (2 Gy or 6 Gy) at different time points (60’) and (45’, 60’) respectively. Lysate of normal splenic wildtype B cells stimulated with PMA (30ng/ml) for 60’ was used as a positive control for EGR1 expression [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0169767#pone.0169767.ref041" target="_blank">41</a>]. RNA extracted from wildtype BM-MNCs in (C) was used to determine EGR1 mRNA expression by qRT-PCR.