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. Author manuscript; available in PMC: 2010 Jan 1.
Published in final edited form as: Free Radic Biol Med. 2008 Oct 7;46(1):33–41. doi: 10.1016/j.freeradbiomed.2008.09.022

Figure 2. Mutation of nuclear localization sequence disrupts p21 nuclear functions but maintains protection during hyperoxia.

Figure 2

H1299 cells with inducible expression of EGFp21 or EGFp21ΔNLS were cultured in the absence (−) or presence (+) of doxycycline (DOX). (A) Total numbers of cells over time were quantified over time in room air (n=4, *p<0.01). (B) Cells were cultured for 2 days in room air (RA) or hyperoxia (O2) in the absence (−) or presence (+) of doxycycline and cell their progression through cell cycle was analyzed. Results shown are representative of three separate experiments. (C) Cells expressing EGFp21, EGFp21ΔNLS or EGFp27 were cultured in doxycyline, transfected with pNF-κB-LUC and treated with 0.5 ng/mLTNF-α for 12 hrs. Cell extracts were prepared and assayed for luciferase activity and data are expressed as a ratio of firefly/Renilla luciferase activity normalized to untreated cells (n=6, *p<0.001). (D) Cell viability of EGFp21 and EGFp21ΔNLS inducible cells during exposure to hyperoxia for 0, 2 and 4 days was assessed on a flow cytometer by gating for propidium iodide positive cells (n=4, *p<0.03).