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. 2011 Sep 5;194(5):689–703. doi: 10.1083/jcb.201105143

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© 2011 Boyd et al.

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Figure 4. — The presence of nucleoli determines p53 levels in micronuclei. (A–D) Examples of U2Os–L11-EGFP cells micronucleated by incubation with 0.1 µg/ml colcemid for 48 h either untreated (A, C, and D) or UV irradiated 5 h before fixation (B) followed by staining for p53 (A and B, red), MDM2 p53 (C, red), or p21 p53 (D, red) and DAPI (blue). (E) Relative expression intensities in a series of cells treated and imaged as in A–D. The normalization parameter for each micronucleated cell was the average of the mean intensity for the stained protein among all nucleolated (Yes) micronuclei. Untreated and UV-irradiated cells were analyzed separately. Plotted values are ratios, and therefore, no direct comparison can be made between the values for the untreated compared with the UV-irradiated cells. Means (closed circles) ± SD are indicated. P-values correspond to t tests for expression differences (null hypothesis: mean protein staining levels are equal between nucleolated and nonnucleolated micronuclei).