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. Author manuscript; available in PMC: 2018 Apr 20.
Published in final edited form as: Cell Chem Biol. 2017 Apr 6;24(4):481–492.e5. doi: 10.1016/j.chembiol.2017.03.008

Figure 4.

Figure 4

Eg5 is deacetylated by HDAC1. A) Chemically acetylated full length recombinant Eg5 was incubated in the absence or presence of recombinant HDAC1 and SAHA for 2 hr at 37°C and analyzed by SDS-PAGE and western blotting with acetyl lysine (top) and a combination of Eg5 and HDAC1 (bottom) antibodies. B) Quantification of the Ac-Lys western blot from part A. Mean and standard error of three independent trials are shown (Figures S8A–D). Quantification of the Eg5 blot as a load control is shown in Figures S8E–F. ***p<0.0001. C) Eg5 was immunoprecipitated from T-Ag Jurkat cells (lane 1) and incubated with immunoprecipitated HDAC1 (lane 2) in the presence or absence of 3 mM SAHA (lane 3) for 1 hr at 30°C, followed by SDS-PAGE separation and immunoblotting with acetyl lysine (top), Eg5 (middle), and HDAC1 (bottom) antibodies. D) Quantification of the Ac-Lys western blot from part C. Mean and standard error of three independent trials are shown (Figures S9A–D). Quantification of the Eg5 blot as a load control is shown in Figures S9E–F. ***p<0.0001