Laminb1

For Western blotting, liver tissue samples (100 mg) were added to RIPA cell lysate (containing 1mM PMSF) and lysed on ice for 30 min. The supernatant obtained after centrifugation at 12,000 rpm for 10 minutes contained the total tissue protein. The extraction of liver nuclear proteins was performed based on previous studies.34 (link) Then, 3× SDS-PAGE protein loading buffer was added to the collected protein samples at 1:2, and the solution was heated in a boiling water bath for 10 minutes to fully denature the protein. After cooling to room temperature, the protein sample was directly loaded on to a SDS-PAGE gel. The samples were then subjected to electrophoresis, membrane transfer, and sealing. For total proteins, the membrane was incubated overnight with the following antibodies: TGR5 (1:5000 dilution, Abcam72608), p-IκBα (1:1000 dilution, Bioss, cat. no. 5515R), IκBα (1:2000 dilution, Abcam, catalog cat. no. 32518), and β-actin (1:1000 dilution, Abcam, cat. no. 8227). For the nucleoprotein, the membrane was incubated overnight with antibodies p-p65 (1:1000 dilution, Santa Cruz, cat. no. 136548), p-p50 (1:1000 dilution, Bioworld, cat. no. 4132), and LaminB1 (1:500 dilution, Bioss, cat. no. 33040M). Finally, an ECL luminescence kit was used to detect signals according to the manufacturer’s instructions.