Figure 3. MWCNTs induce an oxidative stress-dependent nuclear translocation of Nrf2.

THP-1 differentiated into macrophages cells were treated with 1 μg/cm² of MWCNT or Fe@MWCNT suspension in presence or absence of NAC. After 24 h exposure, (A) immunofluorescence was performed to analyze Nrf2 nuclear translocation (scale bars correspond to 10 μm). (B) Nrf2 and Keap1 mRNA quantifications by qPCR were assessed to identify transcriptional regulation. (C) Western blotting on nuclear protein was performed to analyze Nrf2 nuclear translocation. Lamin A was used as nuclear marker. Experiments were repeated at least three times with similar observations in each experiment. Nrf2 was only translocated to the nucleus of cells exposed to MWCNTs. This phenomenon is inhibited by NAC suggesting the important role of oxidative stress in the nuclear translocation of Nrf2. To improve the clarity, gels were cropped but full-length blots are presented in Supplementary Figure S1.