Fig. 6
From: An NF-κB/OVOL2 circuit regulates glucose import and cell survival in non-small cell lung cancer
OVOL2 protein expression is downregulated by P65 in NSCLC cells. A NCI-H661 cells were transduced with empty lentiviral vectors or lentiviral P65 expression vectors. The protein levels of P65 and OVOL2 were evaluated by Western blot analysis (left), and the mRNA expression levels of OVOL2 and OVOL2 target genes (ZEB1, Skp2 and Notch1) were measured by qPCR (right). B SK-MES-1 cells stably expressing Ctrl shRNA or one of two different P65 shRNAs were generated. The protein levels of P65 and OVOL2 were evaluated by Western blot analysis (left), and the mRNA levels of OVOL2, ZEB1, Skp2 and Notch1 were measured by qPCR (right). C NCI-H661 cells were cotransfected with Flag-tagged OVOL2 and HA-tagged P65 or Ctrl vector, and a CHX chase assay was conducted. D NCI-H661 cells were cotransfected with Nluc-OVOL2 and pLV-P65 or Ctrl vector, and a NanoLuc luciferase assay was conducted to assess protein stability. E NCI-H661 cells were co-transfected with Flag-tagged OVOL2 and HA-tagged P65 or Ctrl vector. The cells were then treated with PBS (Ctrl), the proteasome inhibitor MG132 or the autophagy inhibitor CQ. OVOL2 protein stability was evaluated by Western blot analysis. In A, B, and D, the data are expressed as the means ± SDs of three independent experiments. **, p < 0.01; ***, p < 0.001