2.11 Western blot analysis
Proteins were extracted from different treatment groups of M2
macrophages. Protein concentration was determined using an Enhanced BCA
Protein Kit (Beyotime Biotechnology, China). Cell extracts containing
equal amounts of total protein were subjected to 10% sodium dodecyl
sulfate-polyacrylamide gel electrophoresis, and the resolved proteins
were transferred to polyvinylidene fluoride membranes (Invitrogen).
After blocking with 3% BSA for 2 h at room temperature, the membranes
were incubated with primary antibody
overnight at 4 °C. The next day, the membrane was incubated with the
corresponding secondary antibody for 2 h. Finally, an enhanced
chemiluminescence kit was used to detect immunoreactivity.