FIGURE 6.

NAXE inhibits HIF‐1α signaling by eliminating ROS in HCC. A, Representative images show ROS level in indicated cells. DCFH‐DA is used as a probe to detect ROS level. The nuclei are stained by DAPI. B, mRNA and protein expression of indicated molecules in HepG2^shNAXE, Huh7^NAXE, and their control cells with either presence or absence of NAC (10 mmol/L) or H₂O₂ (5 μmol/L) treatment. **P < .01. C, IF analysis shows expression of E‐cadherin and vimentin in NAXE‐interfered cells and their control cells after treated with H₂O₂ (5 μmol/L) or NAC (10 mmol/L) or mock treatment. D, Wound‐heal and transwell assays respectively show the influence of H₂O₂ (5 μmol/L) or NAC (10 mmol/L) on migration and invasion capacity of HepG2^shNAXE, Huh7^NAXE, and their control cells. The data are quantified in Figure S8C,D. E, Mice bearing orthotopic tumors derived from HepG2^shNAXE and Huh7^control cells are treated with NAC by adding into drinking water (supplement with 40 mmol/L NAC for the entire duration of experiment), which could reverse HCC growth and metastasis caused by NAXE downregulation. F, Representative mice lung tissue sections from HepG2^shNAXE and Huh7^control cells treated with or without NAC. The incidence rate of lung metastasis is shown in right bar graphs