Fig. 3.

Histone H2BK120 acetylation by PEG-LANA-DSH in living cells. (A) Structures of PEG-LANA-DSSMe (11–13). (B) LC-MS/MS analysis of acetylation yield for H2BK120 on a recombinant nucleosome (0.37 µM) treated with 3 (10 mM) and TCEP (0.1 mM) in the presence of 1, 2, or 11–13 (2 µM) or in the absence of any catalysts (control) at 37 °C for 5 h in a test tube. Error bars represent the data range of two independent experiments. (C) Representative dot plot of rhodamine signals (horizontal) and cell size (vertical). HeLa S3 cells were loaded with PEG-LANA-DSSMe and dextran-rhodamine, and analyzed by flow cytometry. Judging from the strength of rhodamine signals, high, mid, and low fractions were defined and separated. (D–F) H2BK120 acetylation yield in living cells determined by LC-MS/MS analysis. HeLa S3 cells pretreated with BSO (100 µM) were loaded with 11, 12, or 13 (500 µM), TCEP (2 mM), dextran-rhodamine (0.5 mg/mL), and/or 3 (10 mM), and incubated with growth medium containing 3 (30 mM) at 37 °C for 8 h or indicated time. Acetylation yield of high, mid, or low fractions are shown. Control indicates acetylation yield without any treatment. Error bars represent the data range of two independent experiments.