Enhanced chemiluminescence solution

According to RIPA lysis buffer (Solarbio), total proteins were prepared. Next, 30‐μg protein samples were size‐fractionated by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. After transferring onto PVDF membranes (2 h, 100 V), primary antibodies (4°C, overnight) were incubated. After incubation with secondary antibody, protein bands were observed according to the enhanced chemiluminescence solution (Vazyme, Nanjing, China). Antibodies (Abcam, Cambridge, UK) contains E‐cadherin (ab231303; 1:1500), N‐cadherin (ab98952; 1:1000), HOXB7 (ab152454; 1:1500), β‐actin (ab8227; 1:2000), and secondary antibodies (ab205718/ ab205719; 1:4000). Western blot assay was conducted at least three times.

RIPA lysis buffer (Beyotime) was used for cell lysis and to extract proteins, and the proteins were denatured via heating for 3–5 min at 100°C. Protein samples (about 30 μg/lane) were loaded onto SDS‐PAGE for separating the proteins after quantification of total proteins and then blotted onto PVDF membranes (Millipore). After sealing with 5% nonfat milk (Beyotime), the blocked membrane was immunoblotted for 12–14 h at 4°C using primary antibodies, followed by incubation for 1–2 h with a goat anti‐rabbit IgG secondary antibody (ab205718; 1:4000; Abcam). Lastly, the enhanced chemiluminescence solution (Vazyme) was applied to observe the combined signals. The antibodies including KIAA0101 (ab226255; 1:1000) and β‐actin (ab8227; 1:2000) were bought from Abcam, while E‐cadherin (20874‐1‐AP; 2000) and N‐cadherin (22018‐1‐AP; 1:2000) were purchased from Proteintech.