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. 2011 Aug 31;286(42):36396–36403. doi: 10.1074/jbc.M111.257071

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© 2011 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 4. — p16^INK4a-depleted PRE cells develop early IL-6 secretion with 53BP1 foci formation. A, IL-6 secretion as a function of passage. CM from WI-38 cells infected with control (insertless) or shp16 lentivirus were analyzed for IL-6 by ELISA. We compared early (early PRE) and late (late PRE) passage cells with the same cells at complete replicative senescence (SEN(REP)). B, single cell analysis of p16^INK4a and 53BP1 foci. p16^INK4a and 53BP1 foci were detected by immunofluorescence. Upper panels show p16^INK4a (green) and 53BP1 (red) co-staining. Middle and bottom panels show the grayscale for 53BP1 staining. The yellow arrows indicate cells with three or more 53BP1 foci. C, cell populations shown in B were analyzed for the percentage of nuclei positive for three or more 53BP1 foci. At least 200 nuclei were scored per condition.