Figure 4. Fe@MWCNTs induce an oxidative stress dependent nuclear translocation of Bach1.

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 Bach1 nuclear translocation (scale bars correspond to 10 μm). (B) Bach1 mRNA quantification by qPCR was assessed to identify transcriptional regulation. (C) Western blotting on nuclear protein was performed to analyze Bach1 nuclear translocation. Lamin A was used as nuclear marker. Experiments were repeated at least three times with similar observations in each experiment. Bach1 was only translocated to the nucleus of cells exposed to Fe@MWCNTs. This phenomenon is inhibited by NAC suggesting the important role of oxidative stress in the nuclear translocation of Bach1. To improve the clarity, gels were cropped but full-length blots are presented in Supplementary Figure S1.