Figure 5. Suppression of NRF2 enhances ferroptosis in vivo.

(A, B) NRF2 knockdown Hepa1–6 cells were more sensitive to erastin and sorafenib in vivo. (A) C57BL/6 mice were injected subcutaneously with indicated Hepa1–6 cells (1 × 10⁶ cells/mouse) and treated with erastin (30 mg/kg i.p., twice every other day) and sorafenib (10 mg/kg i.p., once every other day) at day seven for two weeks. Tumor volume was calculated weekly. Data represents mean ± SE (n=5–8 mice/group, * p < 0.05). (B) Q-PCR analysis of the indicated gene expression in isolated tumor at day 28. (C, D) The alkaloid trigonelline increased the anticancer activity of erastin and sorafenib in vivo. (C) C57BL/6 mice were injected subcutaneously with indicated Hepa1–6 cells (1 × 10⁶ cells/mouse) and treated with erastin (30 mg/kg i.p., twice every other day) and sorafenib (10 mg/kg i.p., once every other day) with or without the alkaloid trigonelline (1 mg/kg i.p., once every other day) at day seven for two weeks. Tumor volume was calculated weekly. Data represent mean ± SE (n=5–8 mice/group, * p < 0.05). (D) Q-PCR analysis of the indicated gene expression in isolated tumor at day 28.