2.4 Western blotting
The collected and measured proteins were separated by SDS-PAGE electrophoresis and then electro-transferred onto a PVDF membrane. After blocking with non-fat milk, the membrane was incubated with different primary antibodies at 4°C overnight, respectively, including rabbit anti-TLR2, TLR4, TLR8(MDL,USA);Then, HRP-conjugated secondary antibody was incubated with PVDF membrane, and a chemiluminescence ECL system(Bio-rad,USA) was utilized to detect the immunoreactive proteins.