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FIGURE 3 Carbofuran affects Nrf2 signalling. (A) Schematic illustration of the Nrf2-ARE signalling pathway as well as monitoring the pathway using a transgenic model. Nrf2 translocates into the nucleus by releasing Keap1. In the nucleus, Nrf2 transactivates target genes including gstp1 through binding to ARE regulatory element. The Nrf2-ARE signalling pathway was monitored using a double-transgenic zebrafish line expressing Neh2 domain (of Nrf2)-mCherry and gstp1-EGFP. (B) Effect of carbofuran on the Nrf2-ARE signalling pathway. Embryos were treated with carbofuran for 24 hours, started from 56 hpf and observed under both bright field (BF) (top row) and fluorescence conditions (bottom two rows) of the microscope. Both Neh2-mCherry and gstp1-EGFP expressions were down-regulated by carbofuran exposure. Scale bar: 250 μm. The number of animals was 10 (n = 10). Data are presented as mean ± SE. **P ≤ .01; ***P ≤ .005