Lamin b

Western blot was performed as previously described [21 (link)]. Mice were sacrificed after being anesthetized, and the hippocampi were collected and homogenized with ice cold RIPA lysis buffer. After centrifuging at 12,000 g for 15 min at 4°C, the supernatants were collected, and the protein concentration was measured using a BCA protein assay kit (Bioworld, USA). Equal proteins were separated by 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis gel, transferred onto a PVDF membrane, and incubated in 5% non-fat milk for 2 h at room temperature (RT). Then, the membrane was incubated at 4°C overnight with COX-2 (1 : 2,000, Bioworld, USA), iNOS (1 : 1,000, BD Biosciences, USA), p-IκB (1 : 500, Cell Signaling Technology, USA), IκB (1 : 500, Cell Signaling Technology, USA), p65 (1 : 500, Cell Signaling Technology, USA), GAPDH (1 : 5,000, Bioworld, USA), and lamin B (1 : 2,000, Bioworld, USA). The membrane was washed with TBST for 30 min, and incubated in the proper secondary antibody for 1 h. Then, membranes were washed for 45 min with TBST, and processed with chemiluminescence reagents using an ECL kit (Thermo Fisher, USA). ImageJ (NIH, USA) was used to examine the intensities of the bands.