In vivo Study of the Histone Chaperone Activity of Nucleolin by FRAP

Figure 2

FRAP experiments on H2B-eGFP, H4-eGFP, and macroH2A-eGFP in nucleus of control untransfected cells. (a) Example of photobleaching on H2B-eGFP and nucleolin-mCherry stable cell line. Scale bars represent 5 μm. (b) Normalized fluorescence recovery (%) of H2B-eGFP (green, 6 cells), H4-eGFP (orange, 5 cells) and macroH2A-eGFP (blue, 3 cells) in the nucleoplasm compartment. Each curve represents the fluorescence recovery of a single cell. (c) Average of the normalized fluorescence curves in (b) H2B-eGFP (green), H4-eGFP (orange) and macroH2A-eGFP (blue). (d) Comparison of the percentage of fluorescence recovery at 1h30 for H2B-eGFP (green), H4-eGFP (orange) and macroH2A-eGFP (blue). (e) Dissociation rate constant values 𝑘_off (s⁻¹) of H2B-eGFP, H4-eGFP, and macroH2A-eGFP.

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