Fig. 5

Ethanol metabolism and ROS are required for secretory pathway stress. (A) Heatmap of qPCR data from liver cDNA measuring the expression of genes involved in the UPR. The fold changes of the median C_T values are in supplementary material Table S3. All six clutches are aligned according to the order of the lowest to the highest expression of bip in 350 mM ethanol treatment. (B) Representative images of the tail of Tg(l-fabp:Dbp-EGFP) 120 hpf larvae treated with 0 or 350 mM ethanol or co-treated with AA and 350 mM ethanol for 24 hours. The rectangles in the left panel are magnified in the right panel. Scale bars: 0.2 mm for the left panel and 0.04 mm for the right panel. Note that the curved tail in untreated larvae is an artifact of fixation. (C) Immunoblots of transgenic Tg(l-fabp:Dbp-EGFP) larvae treated as in B using anti-GFP antibody and a non-specific band serving as a loading control. (D,E) Low concentration of ethanol and H₂O₂ synergize to induce UPR (D) and oxidative stress (E). Heatmaps of qPCR data from the livers of larvae exposed to individual treatments of 100 mM ethanol or 2.1 mM H₂O₂, or a co-treatment of the two, for 24 hours. The individual C_T values are in supplementary material Tables S4 and S5 for D and E, respectively. All six clutches are aligned according to the order of the lowest to the highest expression of bip (D) or sod2 (E) in the co-treatment of 350 mM ethanol and 2.1 mM H₂O₂.