Fig. 7

ATP6V1A ubiquitination by FBXO9 suppresses Wnt signaling and EMT A Immunoblot analysis of active β-catenin (non-phospho-β-Catenin in Ser45) levels and other indicated proteins in A549 cells transfected with siCtrl or siFBXO9. GAPDH was used as a loading control throughout. B Immunoblot analysis showing the levels of active β-catenin (non-phospho-β-catenin in Ser45) and indicated proteins in 889DTC cells. The cells were either Fbxo9-knockout or Fbxo9-knockout cells reintroduced with full-length Fbxo9 or Fbxo9-ΔF. C Immunoblot analysis illustrating the levels of active β-catenin and indicated proteins in 889DTC cells with Fbxo9-knockout or Fbxo9-knockout 889DTC cells treated with 200 nM bafilomycin A1 (BAF) for 12 h. D, E Immunoblot analysis illustrating the expression levels of active β-catenin and EMT markers in A549 (d) or H1299 (e) cells ectopically expressing V1A-23 aa. F Experimental setup for tail vein lung metastasis treatment. Nude mice were injected with 1 × 10⁵ 889DTC-LUC cells by vein tail route and received intraperitoneal treatment with a vehicle (5% DMSO in corn oil), BAF (1 mg/kg), lansoprazole (LAN) at 20 mg/kg, or LAN at 40 mg/kg (n = 5–6 mice per group). The treatments were administered every other day for a total of five consecutive treatments over a 21-day period. G, H Lungs of mice were collected on day 21 for both bioluminescent imaging (G) and histological examination using H&E staining (H). The arrows in the image point towards the location of the tumor within the lungs. Scale bar, 100 μm. I Table displaying the incidence of metastatic nodules following different treatments. J Quantitative evaluation of lung metastatic nodules. Data are expressed as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001