Ne per cytoplasmic extraction reagent kit

HDAC inhibitors, tacedinaline (CI994) for HDAC1, RGFP966 for HDAC3, and PCI-34,051 for HDAC were purchased from Selleck Chemicals (Houston, TX, USA). The proteins were extracted with a NE-PER cytoplasmic extraction reagent kit (Thermo Fisher Scientific, Waltham, Massachusetts, USA) for western blotting. The protease inhibitor was purchased from Millipore (Billerica, Massachusetts, USA). SV40 LT mouse monoclonal antibody, which cross-reacts with BKPyV LT, was purchased from Millipore. Primary antibodies against HDAC1, HDAC2, HDAC3, and HDAC8 were purchased from Cell Signaling Technology Inc. (Danvers, Massachusetts, USA). GAPDH expression was used as an internal control for western blotting, and the GAPDH antibody (monoclonal, rabbit anti-human GAPDH) was purchased from Cell Signaling Technology Inc.

Nuclear extraction was conducted using a NE-PER cytoplasmic extraction reagent kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. Briefly, the cytoplasmic fraction was extracted with cytoplasmic extraction buffers I and II by vortexing. The nuclear fraction was extracted in nuclear extraction buffer, and the protein was quantified using Bio-Rad Protein Assay reagent (Bio-Rad, Hercules, CA, USA).