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. 2020 Jan 23;48(5):2401–2411. doi: 10.1093/nar/gkaa011

Figure 4.

Figure 4.

Lysine-acetylation of the total and the RNAP-associated HrdB proteins at the exponential growth phase. (A) Lysine-acetylation of the total and the RNAP-associated HrdB proteins. Cell lysates (2 mg) prepared from JE04 cells at early exponential phase were subjected either to immunoprecipitation with anti-HrdB antibody to collect total HrdB or to purification of RNAP as described in Materials and Methods. Immunoprecipitate (HrdB-IP) (3 μl for HrdB and 15 μl for AcK) and purified RNAP samples (1 μg for HrdB and 5 μg for AcK) were analyzed by western blot using either anti-HrdB or anti-AcK antibodies. Representative images from seven independent experiments are shown. (B) Relative ratio of HrdB acetylation in total vs. RNAP-bound HrdB. The lysine-acetylation ratio of HrdB in the RNAP was compared with that of the total immunoprecipitated HrdB, which was set to 1.0. Quantification from seven independent experiments is presented with an average value and standard error. Statistical significance with a P value of <0.001 (***) is indicated.