Skip to main content
. Author manuscript; available in PMC: 2018 Aug 9.
Published in final edited form as: Cell Metab. 2018 Jun 7;28(2):228–242.e6. doi: 10.1016/j.cmet.2018.05.012

Figure 6. p22phox Plays an Important Role in ROS Generation in mel + TNF-α-Treated Tumor Cells.

Figure 6

(A) Mitochondria are not the major source of ROS production in mel + TNF-α-treated tumor cells. CT26HA cells were treated as indicated. Cells were evaluated for ROS and superoxide levels by CM-H2DCFDA and mitoSOX staining, respectively.

(B) p22phox-deficient tumor cells are less sensitive to the toxicity of mel + TNF-α. CT26HA.ΔCyba and the parental CT26HA cells were subjected to the indicated culturing conditions. After overnight incubation, cell viability was evaluated by DAPI staining. Error bars indicate SEM.

(C) CT26HA.ΔCyba cells exhibit reduced ROS levels after mel + TNF-α treatment. CT26HA.Δ Cyba and CT26HA cells were treated with mel + TNF-α overnight before evaluating ROS levels by CM-H2DCFDA.

(D and E) (D) Treatment-induced GSH reduction is alleviated in CT26HA.ΔCyba cells. CT26HA.ΔCyba and CT26HA cells were either untreated or treated with mel + TNF-α for 7 hr before being harvested for GSH and GSSG measurements. Percentage GSH reduction, calculated using the formula (GSH in untreated cells – GSH in treated cells)/GSH in untreated cells, is summarized in bar graph and shown as mean ± SEM. Changes in GSH/GSSG ratio are shown in (E).

**p < 0.01, ***p < 0.001. See also Figure S6.