Comprehensive Protein Analysis of EMT

Total protein was isolated from cells using RIPA lysis buffer (Beyotime). Western blot analysis was performed as previously described 15 (link). The intensity of the protein bands was quantified using ImageJ software (NIH). Primary antibody against FN1 (1:1000, ab32419) was purchased from Abcam (Shanghai, China). Antibodies against THBS1 (1:1000, 18304-1-AP), CPED1 (1:1000, 20924-1-AP), AMOTL2 (1:1000, 23351-1-AP), TGFβ1 (1:1000, 21898-1-AP), TGFβ2 (1:1000, 19999-1-AP), and GAPDH (1:10000, 60004-1-Ig) were purchased from Proteintech Group (IL, USA). Antibodies against E-cadherin (1:500, #3195), N-cadherin (1:500, #13116), Slug (1:1000, #9585), Snail (1:1000, #3879), ZO1 (1:1000, #8193), β-catenin (1:1000, #8480), CLDN1 (1:1000, #13255), MMP2 (1:1000, #40994), and MMP9 (1:1000, #13667) were purchased from Cell Signaling Technology (MA, USA).