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. 2014 Aug 18;10(8):746. doi: 10.15252/msb.20145377

Figure 6. Propagation of H3K9me3 by stably tethered SUV39H1 to nucleosomes in its spatial proximity.

Figure 6

  1. Establishment of H3K9me3 domains by GFP-SUV39H1 recruited to the nuclear lamina via GBP-Lamin B1 in iMEF Suv39h dn cells. H3K9me3 was detected via CD-RFP and appeared in a confined region adjacent to the nuclear lamina (see inset). When the inactive mutant SUV39H1-H324L-GFP was recruited, no enrichment of CD-RFP was observed. Scale bar, 10 μm.
  2. Cartoon model depicting the experimental setup in panel A.
  3. Averaged radial fluorescence intensity profiles from the lamina to the center of the nucleus measured for the experiments described in panel A. The profile of CD-RFP reflects the H3K9me3 levels and was measured in cells transfected with GFP-SUV39H1 (red) or the inactive SUV39H1-H324L-GFP mutant (blue, control). The recruitment of GFP-SUV39H1 (green) resulted in a lamina-confined enrichment with a width of 0.40 ± 0.02 μm as determined by fitting the data to an exponential decay curve. While wild-type SUV39H1 methylated the surrounding chromatin within a confined area of 0.47 ± 0.03 μm width (red), SUV39H1-H324L (blue) did not increase the methylation (blue) in this region. Error bars correspond to SEM.