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. 2010 May 24;107(23):10454–10459. doi: 10.1073/pnas.1003064107

Fig. 3.

Fig. 3.

FRAP. HeLa cells expressing GFP-H3.1 or GFP-H3T were subjected to the FRAP analysis. (A) The mobility of GFP-H3.1 or GFP-H3T in living cells was analyzed by bleaching one-half of the nucleus. (B) The averages of the relative fluorescence intensity of bleached area were plotted with the standard deviations (n = 5). (C) GFP-H3T was incorporated into the HeLa cell chromatin. (Upper ) DNA fragments of mono-, di-, and trinucleosomes fractionated by sucrose gradient centrifugation were analyzed by agarose gel electrophoresis with ethidium bromide staining. (Right and Left) The nucleosome samples from the HeLa cells with and without GFP-H3T expression, respectively. The sucrose gradient fraction numbers are indicated at the top of each panel. Middle panel. Histone compositions of the purified nucleosomes were analyzed by 16% SDS-PAGE with Coomassie brilliant blue staining staining. (Lower) GFP-H3T was detected with anti-GFP monoclonal antibody.