FIGURE 2.

Yes‐associated protein (YAP) positively regulates AXL expression in head and neck squamous cell carcinoma (HNSCC) cells. A, YAP and AXL protein expression were detected in HNSCC cell lines and normal oral mucosal epithelial cell. B, The expression of AXL and YAP was determined after transfection with si‐YAP for 48 h. C, The percentage of AXL‐ and YAP‐positive cells was determined using immunofluorescence transfected with si‐YAP for 24 h. Scale bars: 50 μm. D, RNA sequencing analysis of mRNA extracted after transfection with si‐YAP for 48 h. Significantly altered genes associated with regulation of Hippo signaling and tyrosine kinase activity and corresponding log₂ fold change are listed in the heat map. (blue, downregulation; red, upregulation). E, AXL expression was detected after YAP inhibitor verteporfin for 24 h. F, AXL expression was determined after ectopic expression of YAP for 48 h. G, The main downstream molecules of AXL were detected after treatment with 1 μmol/L verteporfin for 24 h. H, The binding site of TEAD1 in AXL‐promoter was predicted and mutant AXL‐promoter was designed. I, Wild‐type and mutant AXL‐promoter activities were detected after cotransfection with YAP plasmid for 24 h. *P < .05, **P < .01, ***P < .001, based on Student t test