Nusap1

Western blotting analysis was conducted using NUSAP1 (1:1000; #12024-1-AP; Proteintech, Rosemont, IL, USA), YAP1 (1:1500; #14074; Cell Signaling Technology, Danvers, USA), LATS1 (1:1500; #3477; Cell Signaling Technology, Danvers, USA), LATS2 (1:2000; #5888; Cell Signaling Technology, Danvers, USA), CTGF (1:2000; #86641; Cell Signaling Technology, Danvers, USA), CYR61 (1:2000; #14479; Cell Signaling Technology, Danvers, USA), anti-HA (1:2500; #ab9110; Abcam, Cambridge, USA), anti-Flag (1:2500; #A2220; Sigma-Aldrich, USA), β-actin (1:3000; #AF7018; Affinity, Jiangsu, China) and GAPDH (1:3000; #A2220; Affinity, Jiangsu, China). Human GC samples originally obtained from the First Affiliated Hospital of Nanchang University, along with the xenograft tumors, were ground and lysed in lysis buffer before Western blotting analysis. The Western blotting experiments were performed as previously described (28 (link)). GAPDH or β-actin was used as an internal control, and all the protein bands were analyzed using ImageJ software.

Western blotting was performed as described in our previous study [17 (link), 18 (link)]. The antibodies used in the present study were against NUSAP1 (1:3000; Proteintech), GAPDH (1:5000; Abcam), E-cadherin (1:1000; CST), ZEB1 (1:1000; CST), Claudin 1 (1:1000; CST), Snail (1:1000; CST), Zo 1 (1:1000; CST), AMPKα (1:1000; CST) and Phospho-AMPKα (1:1000; CST). The membranes of western blotting were cut to a suitable size prior to binding with antibodies.