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. 2008 Feb 7;36(5):1713–1722. doi: 10.1093/nar/gkn014

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© 2008 The Author(s)

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Loss of p21 enhances chromatin-binding and association between p300 and PCNA after DNA damage. (A) Western blot analysis of recruitment of p300, PCNA and p21 proteins in the chromatin-bound fraction. Human h-TERT–immortalized p21^+/+ and p21^−/− fibroblasts were irradiated with UV-C (10 J/m²), collected at the indicated time points (h) after UV irradiation, and fractionated as described in ‘Materials and Methods’ section. Actin was also determined as a loading control. (B) Western blot analysis of proteins remaining chromatin-bound 24 h after UV-irradiation in LF1 fibroblasts not transfected, or transfected with p21-specific or control (GFP) siRNA pools. (C) Immunoprecipitation (Ip) with anti-p300 or irrelevant antibody (Ig) was performed on extracts of chromatin-bound proteins from p21^+/+ and p21^−/− fibroblasts collected 0.5 or 24 h after UV irradiation. Input load: 1/15 of nuclear extracts.