TABLE 1.

Fluorescence anisotropy of Cy3 and Cy5 labeled at the 601A and 601B nucleosomes

Anisotropy was measured with the excitation wavelength of 532 nm (for Cy3) or 600 nm (for Cy5) and emission wavelength of 570 nm (for Cy3) or 670 nm (for Cy5). For FRET anisotropy (i.e. Cy5 emission anisotropy when it was excited via FRET) was measured at 670-nm emission with 532-nm excitation. The wavelength range was ±2.5 nm for excitations (Ex) and ±14 nm for emissions (Em). β is the average angle between the emission dipole of Cy3 and the absorption dipole of Cy5 (see the supplemental materials). β was calculated based on the measured anisotropy (r) using the formula r = 2/5{(3cos² β − 1)/2}. The detailed calculation procedure is described in the supplemental materials. CTL-NUC, unacetylated nucleosomes; CTL-PCL, unacetylated nucleosomes incubated with Piccolo NuA4 complex in the absence of acetyl-CoA; PCL, acetylated nucleosomes incubated with Piccolo NuA4 complex in the presence of acetyl-CoA.

Ex/Em	601A				601B
	532/570 (Cy3)	600/670 (Cy5)	532/670 (Cy5 via FRET)	β	532/570 (Cy3)	600/670 (Cy5)	532/670 (Cy5 via FRET)	β
Free	0.200 ± 0.003	0.130 ± 0.004			0.200 ± 0.003	0.130 ± 0.004
CTL-NUC	0.314 ± 0.011	0.329 ± 0.009	0.140 ± 0.011	37.8 ± 1.0	0.338 ± 0.010	0.328 ± 0.015	0.078 ± 0.008	45.7 ± 0.5
CTL-PCL	0.321 ± 0.008	0.343 ± 0.013	0.144 ± 0.011	37.9 ± 1.0	0.337 ± 0.014	0.337 ± 0.013	0.078 ± 0.011	45.7 ± 0.9
PCL	0.319 ± 0.010	0.347 ± 0.012	0.105 ± 0.010	42.4 ± 0.9	0.347 ± 0.015	0.331 ± 0.014	0.078 ± 0.012	45.8 ± 1.0