Fig. 4
JAK1 interacts with HIF-1/2α and reduces HIF-1/2α protein levels. A, B RT-qPCR of the indicated mRNAs in WCLs from parental and JAK1-knockdown cells (KLE and SPEC-2 cells). All the cells were incubated under hypoxia (2% O2) for 4 h before harvesting. Data are shown as means ± SD (n = 3). C, D Western blotting of the indicated proteins in WCLs from parental and JAK1-knockdown cells (KLE and SPEC-2 cells). The cells were incubated under hypoxia (2% O2). After hypoxia for 0, 2 h, 4 h, cells were harvested, respectively. E, F Western blotting of the indicated proteins. The cells (KLE and SPEC-2 cells) were treated with DMSO or Ruxolitinib (1 μmol) for 48 h. Then cells were incubated under hypoxia (2% O2). After hypoxia for 0, 2 h, 4 h, cells were harvested, respectively. G Western blotting of the indicated proteins in WCLs from 293 T cells co-transfected with Flag-tagged HIF-1/2α expression plasmid and different concentrations of Myc-tagged JAK1 expression plasmid (200 ng, 400 ng, 800 ng, 1200 ng). H Co-IP assays in KLE cells incubated under hypoxia (2% O2) for 24 h. I. Co-IP assays in 293 T cells transfected with the indicated plasmids