Fig. 6

Potential of DEPM to induce nuclear translocation of Nrf2 in endothelial cells after different incubation times. Tetracultures were exposed to 240 ng/cm² of DEPM. Nrf2 translocation was evaluated for cells indirectly exposed to DEPM at 3, 4 and 5 h after exposure. Cells exposed for the same time but without particles in the air stream were used as controls for the fluorescence signals. Cells were fixed and stained for nucleus (Hoechst 33342) and anti-Nrf2 (green). Images were analyzed using a Zeiss LSM 510 META. At 3 h after exposure, first cells showed nuclear translocation. At 4 h after exposure, nuclear translocation was quite obvious in the majority of the cell population. At 5 h after exposure almost every endothelial cells showed strong colocalization of Nrf2 with the nucleus (counterstained in blue). In control EA.hy 926 cells, no nuclear translocation was observed at any time-point (compare with the representative control image)