The cells were extracted with cold RIPA buffer (Beyotime, Beijing, China). The cell lysates were standardized on the basis of protein content. Equal protein lysates (30 μg/lane) were separated by 10% SDS-PAGE and blotted onto polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). The membranes were incubated overnight at 4°C with primary antibodies, including IL-1β (1:1000; Abcam), IL-6 (1:1000; Proteintech, Wuhan, China), IL-10 (1:500; Abcam), TNF-α (1:500; Abcam), TLR2 (1:1000; Abcam), TLR4 (1:200; Santa Cruz Biotechnology), AIF (1:1000; Proteintech), BAD (1:1000; Abcam), BAX (1:1000; Proteintech), P-p65 (1:2000; Abcam), p65 (1:2000; Abcam), IκBα (1:1000; Abcam), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH, 1:12000; Proteintech). The next day, the membranes were incubated with peroxidase-conjugated secondary antibodies (1:5000; Proteintech) for 1 hour at room temperature. An enhanced chemiluminescence substrate kit (Millipore) was used for chemiluminescent detection of reactive signals with autoradiography films (Amersham, Little Chalfont, UK).