DC WB original data

Description

The western blot data which used in "Changing HDAC activity directly reprogrammes murine embryonic stem cells to trophoblast stem cells".

Steps to reproduce

Cells were washed once with PBS and lysed with RIPA lysis buffer (P0013, Beyotime) which containing 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 0.1% SDS, 1 mM EDTA, 1% Triton X-100 and fresh-added 1×protease inhibitor cocktail. The concentration of protein in the samples were detected with the BCA protein assay kit (P0012S, Beyotime). 10 μg proteins of each sample were used as a loading content by 10% SDS-PAGE and transferred sample and marker to PVDF membrane (IPVH00010, Millipore). The membranes were blocked with 5% skim milk powder in TBST for 1.5 h at room temperature and incubated overnight at 4°C with primary antibodies, which are listed in Table S6. After incubation, membranes were washed with TBST for 10 mins (three times), and incubated with secondary antibodies (1:5000; Cell Signaling Technology). Membranes were washed with TBST for 10 min (three times), and the bands were visualized by enhanced chemiluminescence using ECL (P0018FS, Beyotime) and captured on Molecular Imager Gel Doc XR+ (Bio-Rad).

Institutions

Categories

Funding

Licence