Gb13030

MKN28 and AGS gastric cancer cells were lysed using RIPA lysis buffer (Beyotime, Beijing, China) and protease inhibitors (Roche, CA, USA). The proteins were separated by 10% SDS polyacrylamide gel electrophoresis and subsequently transferred onto nitrocellulose (NC) membranes. The membranes were blocked in TBS buffer containing 5% skim milk for 1 h at room temperature. After incubation with primary and secondary antibodies, ECL reagent was used to visualize the protein bands. CLDN6 (ab107059, Abcam), p-LATS1/2 (ab111344, Abcam), LATS1/2 (202761-AP, Proteintech), p-YAP1 (ab76252, Abcam), YAP1 (13584-1-AP 6900-1-Ig, Proteintech), snail1 (13099-1-AP, Proteintech), twist1(25465-1-AP, Proteintech), zeb1 (21544-1-AP, Proteintech), E-cadherin (20874-1-AP, Proteintech), N-cadherin (GB111009, Servicebio), Vimentin (GB11192, Servicebio), Ki67 (GB13030, Servicebio), β-actin (GB11001, Servicebio), HRP (GB23301, GB23303, Servicebio), Lamin B1 (AB0054, Abways).

The apoptosis of transfected OS cells in the xenograft tumors was detected using the TUNEL kit (Roche, Basel, Switzerland), as we previously described. Ki67 was measured using a primary antibody (1:200, GB13030; Servicebio, Wuhan, China).