Fig. 3

Fig. 3 Nrf2 is activated in hepatocytes following liver injury and is required for survival (A) Schematic of the compound transgenic line Tg(lfabp:NLS-mCherry; gstp1:EGFP) used to visualize hepatocyte nuclei and Nrf2 activity. (B) Representative multiphoton images of 5 dpf zebrafish treated with DMSO or APAP for 24 h. Cyan dashed line borders the liver. Scale bars, 50 ?M. (C) Quantification of Nrf2-activated hepatocytes (% GFP+ and mCherry+ cells) acquired from multiphoton imaging of 5 dpf zebrafish treated with DMSO or APAP for 24 h. p value was calculated using an unpaired two-tailed Student?s t test comparing each sample to the respective DMSO control, n = 4. (D) Quantification of Nrf2-activated hepatocytes (% GFP+ and mCherry+ events) as determined by flow cytometry of 5 dpf zebrafish treated with DMSO or APAP for 24 h. p value was calculated using an unpaired two-tailed Student?s t test comparing each sample to the respective DMSO control, n = 3. (E and F) Volcano plots of differentially expressed genes identified by comparing dissected livers of 5 dpf zebrafish treated with DMSO or APAP for 12 (E) and 24 h (F) by RNA-seq analysis. Green points represent Nrf2 target genes. Genes within red area of volcano plots are significantly upregulated (log2FC > 0.6, p < 0.05), and genes within blue area of volcano plots are significantly downregulated (log2FC < 0.6, p < 0.05). (G) Survival data of WT and Nrf2KO zebrafish (n = 120) treated with DMSO or APAP for 72 h from 4 dpf. p value was calculated using a Mantel-Cox test comparing the survival of Nrf2KO APAP to WT APAP. Data are shown as mean ± SEM. See also Figure S3.