Figure 5
From: Osteopontin facilitates tumor metastasis by regulating epithelial–mesenchymal plasticity
iOPN regulates AKT1 expression by interacting with HIF2α. (a) Immunoblot and qPCR analysis of AKT1 in Hep3B cells with knockdown or overexpression of HIF2α. *P<0.05, **P<0.01. (b and c) Dual luciferase reporter assay of AKT1 promoter mutants in collected 293 T cells 48 h after indicated transfection. Each sample was performed in triplicate. Data represent mean±S.D. (d) In Hep3B cells transfected with the indicated lentivirus, chromatin was immunoprecipitated using indicated antibodies, corresponding DNA fragments were then amplified and measured. (e) Confocal microscopy of OPN and HIF2α in 293 T, Hep3B, and HCCLM3 cells. Images are representative of three experiments. Scale bar, 10 μm. (f) lysates from 293 T, Hep3B, and HCCLM3 cells were immunoprecipitated with denoted antibodies. HIF2α and OPN levels were then assessed. (g) Co-immunoprecipitation (Co-IP) analysis of V5-iOPN and HA-HIF2α proteins
