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. 2010 Nov 16;10(2):M110.004317. doi: 10.1074/mcp.M110.004317

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

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Fig. 7. — Preclusion of serine 279 phosphorylation alters HDAC5 localization. A, Cellular localization of stably transfected wild type (WT), S278A/S279A, and S279A HDAC5-EGFP was assessed by direct immunofluorescence microscopy in U2OS cells (bar, 20 μm). Increased cytoplasmic accumulation of HDAC5 (EGFP) was observed because of Ser-to-Ala mutations. Nuclear and HDAC5 (EGFP) boundaries used for the quantification of HDAC5 cellular localization are shown. B, Distribution plot of the percent of HDAC5-EGFP-expressing U2OS cells as a function of the fraction of total HDAC5-EGFP within the cytoplasm (cytoplasmic proportion). Cytoplasmic proportion of HDAC5 was calculated by dividing the integrated intensity of the EGFP signal within the cytoplasm by the total integrated intensity of EGFP within each cell (see Experimental Procedures). WT HDAC5 (red bars; n = 848), S278A/S279A (black bars; n = 868), and S279A (green bars; n = 970) cells showed a median cytoplasmic proportion of 0.20, 0.36, and 0.45, respectively.