D120536

Total protein was extracted and subjected to 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The membranes were first incubated with the primary antibodies against Src (1: 500) (D120536, Sango Biotech, Shanghai, China) Fn14 (1: 500) (D121267, Sango Biotech, Shanghai, China), IκBα (1: 500) (bs-1287R, Bioss, Beijing, China), p-IκBα (1: 500) (bs-18129R, Bioss, Beijing, China), IKKβ (1: 500) (bs-4880R, Bioss, Beijing, China), p-IKKβ (1: 500) (bs-3233R, Bioss, Beijing, China), NF-κB p65 (1: 400) (PB0073, Boster, Wuhan, China), E-cadherin (1: 5000) (No. ab76055, Abcam, Cambridge, UK), N-cadherin (1: 5000) (No. ab18203, Abcam, Cambridge, UK), β-actin (1: 1000) (sc-47778, Santa Cruz, CA, USA) or Histone H3 (1: 500) (bs-17422R, Bioss, Beijing, China) at 4°C overnight and then incubated with HRP-conjugated secondary antibodies (A0208, A0216, A0521, Beyotime, Shanghai, China) for 45 minutes. The integrated ODs (IODs) of the blots were calculated by the Gel-Pro-Analyzer (Media Cybernetics, USA).

Antibodies against Fn14 (D121267) and Src (D120536) were purchased from Sango Biotech (Shanghai, China). Antibodies against IκBα (bs-1287R), phosphorylated IκBα (p-IκBα) (bs-18129R), IKKβ (bs-4880R), p-IKKβ (bs-3233R), and Histone H3 (bs-17422R) were purchased from Bioss (Beijing, China). Antibody against nuclear NF-κB p65 (PB0073) was purchased from Boster (Wuhan, China). Antibody against NF-κB p65 (10745-1-AP) used for immunofluorescent staining was purchased from Proteintech (China). Secondary goat anti-rabbit IgG-HRP (A0208), goat anti-mouse IgG-HRP (A0216), and cy3-labled goat anti-mouse IgG (A0521) antibodies were purchased from Beyotime (Shanghai, China). Antibody against β-actin (sc-47778) was purchased from Santa Cruz (CA, USA). RNA Purified Total RNA Extraction Kit (RP1201) and super M-MLV reverse transcriptase (RP6502) were purchased from BioTeke (Beijing, China). Total Protein Extraction Kit (P0027) was purchased from Beyotime (Shanghai, China). MTT (M-2128) was purchased from Sigma (USA).