Lysates were prepared from cultured RA-FLS as previously described [17 (link)]. Cell nuclear and cytoplasmic extracts were prepared using the Mammalian Nuclear and Cytoplasmic Protein Extraction Kit (TransGen Biotech, Beijing, China) following the manufacturer’s protocol. Protein concentrations were measured using a Quick Start™ Bradford Protein Assay (Bio-Rad, USA).
Western blot assays were performed as previously described [16 ]. Immunoreactive bands were visualized using the Enhanced Chemiluminescence Detection Kit (Invitrogen, USA). Image J software was used to measure the intensity of each band. The relative level of each protein of interest was normalized to the endogenous β-actin, GAPDH or lamin B1 in each experiment.
Western blot assays were performed as previously described [16 ]. Immunoreactive bands were visualized using the Enhanced Chemiluminescence Detection Kit (Invitrogen, USA). Image J software was used to measure the intensity of each band. The relative level of each protein of interest was normalized to the endogenous β-actin, GAPDH or lamin B1 in each experiment.
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