Ab134155

After the total proteins were extracted from cells in each group, the protein concentration was detected using the BCA Protein Assay Kit (Beyotime: P0012), and the samples were added with 5× electrophoresis buffer according to the proportion and placed in a metal bath (100° C) for 10 min. Next, the protein samples were loaded (20 μg/well), separated by SDS-PAGE, and transferred onto a PVDF membrane. After being blocked with 5% skim milk powder (OXOID: LP0031B) or 5% Fatty Acid Free BSA (WarBio: M9203), the membrane was incubated with primary antibodies against cyclinD1 (Abcam: ab16663, 1:200), cyclinA2 (Abcam: ab181591, 1:2,000), proliferating cell nuclear antigen (PCNA) (Abcam: ab29, 1:1,000), matrix metalloproteinase 2 (MMP2) (Abcam: ab92536, 1:1,000), MMP3 (Abcam: ab52915, 1:1,000), MMP9 (Abcam: ab76003, 1:1,000), BDNF (Abcam: ab108319, 1:1,000), phosphorylated tropomyosin kinase receptor B (p-TrkB) (Abcam: ab229908, 1:1,000), total TrkB (t-TrkB) (Abcam: ab134155, 1:1,000), p-ERK1/2 (Abcam: ab184669, 1:10,000), t-ERK1/2 (Abcam: ab17942, 1:1,000) and GAPDH (Abcam: ab8245, 1:5,000) at 4° C overnight. The next day, the membrane was rinsed and incubated with the secondary antibody (Abcam: ab97080, 1:5,000) at room temperature for 1 h. Afterwards, the membrane was rinsed and the proteins were visualized using ECL reagent, followed by statistical analysis.