Figure 1 | Scientific Reports

Figure 1

From: Effects of charge-modifying mutations in histone H2A α3-domain on nucleosome stability assessed by single-pair FRET and MD simulations

Figure 1

Comparison between wild type and mutated nucleosomes with different fluorescent and non-fluorescent based methods. (a) Electrophoretic mobility shift assay with reconstituted nucleosomes analyzed on native 6% PAAG-Gel. Successfully reconstituted nucleosomes bearing mutations in H2A and wild type nucleosomes show decreased electrophoretic mobility in comparison to free DNA. Small differences in electrophoretic mobility between the mutated nucleosomes are detectable suggesting changes of the hydrodynamic radius. (b) Representative AFM pictures of wild type and H2A R81E/R88E nucleosomes reconstituted on a 660 bp DNA containing Widom 601 sequence in the middle of the fragment. Scale bar = 50 nm. (c) Normalized average proximity ratio as a function of NaCl concentration of three independent replicates. Charge-modifying mutations reduced the overall stability of nucleosomes. Black: wild type, Red: H2A R81A mutant, Green: H2A R81E/R88E mutant, Symbols: measured values and standard error of the mean, Lines: sigmoidal fit.

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