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. 2019 Nov 22;111(1):127–136. doi: 10.1111/cas.14224

Figure 3.

Figure 3

Nuclear factor erythroid 2 (NF‐E2)‐related factor 2 (Nrf2) signaling limits cancer cell sensitivity to combination therapy with sulfasalazine (SSZ) and oxyfedrine (OXY). A, Immunoblot analysis of Nrf2, xCT, and β‐actin (loading control) in HCT116, HSC‐4, and A549 cells. B, A549 cells were cultured for 48 h in the absence or presence of SSZ (400 µmol/L), buthionine sulfoximine (BSO; 100 µmol/L), or OXY (50 µmol/L), as indicated, and were then assayed for cell viability. Data are means ± SD from three independent experiments. **P < .01 (Student's t test). C, Immunoblot analysis of Nrf2, xCT and ALDH3A1 in A549 cells transfected with a control of Nrf2 siRNA. D, A549 and A549 transfected with siNrf2 cells were cultured with or without OXY (50 µmol/L) for 48 h, stained with ALDEFLUOR (Stemcell Technologies) in the absence or presence of the aldehyde dehydrogenase (ALDH) inhibitor diethylaminobenzaldehyde (DEAB; 15 µmol/L), and then analyzed by flow cytometry for ALDH activity. SSC‐A, side scatter area. E, A549 cells transfected with control or Nrf2 siRNAs were cultured for 48 h as in (B) and then assayed for cell viability. Data are means ± SD from three independent experiments. **P < .01 (Student's t test). F, A549 cells were cultured for 48 h in the absence or presence of SSZ (400 µmol/L), BSO (100 µmol/L), OXY (50 µmol/L), or ML385 (5 µmol/L) as indicated, and were then assayed for cell viability. Data are means ± SD from three independent experiments. **P < .01 (Student's t test)