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Fig. 3 | Epigenetics & Chromatin

Fig. 3

From: Tissue-specific expression of histone H3 variants diversified after species separation

Fig. 3

Six of 14 novel H3 variants incorporate into chromatin. a Images of GFP-tagged histone H3 variants show localizations in nuclei that were categorized into three groups. Variant expression was induced by tetracycline. Scale bar 10 μm. b Fluorescence recovery after photobleaching experiments to evaluate the nucleosomal stability of H3 variants. The areas marked with white circles were photobleached, and the recovery of the respective fluorescent signals was monitored (selected time points are shown). c GFP-tagged histone H3 variants were categorized into two groups: those that were stably incorporated into nuclei and those that were not. The mobility of the GFP-fused histone H3 variants was analyzed based on fluorescence recovery after photobleaching. Recovery curves of GFP-fused histone H3 variants are shown. Relative fluorescence intensities are mean ± standard deviation (n represents the number of trials). The rate of fluorescence recovery indicates the stability of the chromatin incorporation. Red lines show stable incorporation for GFP-H3.1, H3.2, H3.3, H3t, H3mm7, H3mm11, H3mm12, H3mm13 and H3mm16. Black lines show diffusion in the nucleus for GFP-H3mm6, H3mm8, H3mm9, H3mm10, H3mm14, H3mm15, and H3mm18. Scale bars 10 μm. d Exogenously expressed GFP-tagged histone H3 variants did not interfere with the endogenous expression of core histones. Immunoblots were performed using acid extract from GFP-tagged histone H3 variant-expressing C2C12 cells in the undifferentiated state. The top four panels show exogenously expressed GFP-tagged histone H3 variants; the lower five panels show the endogenous level of each core histone. Wild-type C2C12 cells (WT) were used as a control (no expression of GFP-tagged histones)

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