Fig. 3
ROS production is enhanced in EtBr-treated wild type and djr-1.1 worms, but not hif-1 mutant worms. a In vivo ROS levels were determined in L4/adult worms grown on worm plates containing either EtBr (50 μg/mL) or water. Worms were subsequently incubated in buffer containing H2DCF-DA and whole organism fluorescence intensity measured using a Zeiss LSM 700 confocal microscope. Results were summarized from at least 4 independent experiments. Fluorescence intensity was quantified using ImageJ software. The y-axis depicts absolute values of mean intensity per worm. Statistical analysis was performed in GraphPad Prism 5.0. Asterisk marks denote significant difference between control and drug-treated worm samples for each strain (* p < 0.05, ** p < 0.01, *** p < 0.001). b sod-3 is overexpressed in EtBr-treated wild type and djr-1.1 animals but not EtBr-treated hif-1 animals. mRNA levels of sod-3 was determined by qRT-PCR in wild type, hif-1 and djr-1.1 mutants grown on worm plates containing nematode growth media in either the absence (labeled 0) or presence (labeled 50) of 50 μg/ml of EtBr, using act-1 as a control. Asterisk/plus signs indicate statistical significant difference in expression between control N2 sample and other strains and that between untreated and EtBr-treated sample for each strain (*/+ denotes p < 0.05, **/++ p < 0.01, ***/+++ p < 0.001). sod-3 transcript levels were significantly higher in wild type and djr-1.1 animals, but not hif-1 mutants. c Wild type sod-3::gfp and hif-1;sod-3::gfp strains adults were placed on plates containing EtBr and images were taken of five to 10 worms three and four days post egg-laying for hif-1;sod-3::gfp and wild type animals, respectively