Figure 4.

Effect of 2ME2 and hypoxia on HIF-1α nuclear accumulation in H1N1 infection. Before cells were infected with H1N1 at multiplicity of infection (MOIs) of 1 and 5 for 24 h, A549 and THP-1 cells were pretreated with 0.1 and 0.2 μM 2ME2 for 30 min. A549 and THP-1 cells were exposed to 1% O₂ for 24 h after H1N1 infection. (A and B) Cells were lysed using RIPA buffer to extract the total protein. (C–F) Nuclear and cytoplasmic extraction reagents were used to extract the nuclear and cytoplasmic proteins from the cells. Western blots were performed. β-actin was used as a loading control for cytoplasmic protein and lamin B1 was used as a loading control for nuclear protein. The intensity of the protein bands from three typical experiments was quantified using Image J software. (G, H) Total cellular RNA was extracted and HIF-1α mRNA levels and viral RNA levels were measured by qRT-PCR. β-actin expression was used as an internal control. Data are shown as the mean±s.e.m. of three independent experiments. *P<0.05, **P<0.01 compared with the control group. ^#P<0.05, ^##P<0.01 compared with the H1N1-infected group.