Phthalates Induce Neurotoxicity Affecting Locomotor and Thermotactic Behaviors and AFD Neurons through Oxidative Stress in Caenorhabditis elegans
Figure 5
Effects of phthalates exposure on intracellular reactive oxygen species production in C. elegans.
Synchronized wild-type L1 larvae were incubated in liquid S-basal containing E. coli OP50 bacteria, at 109 cells/mL in the (A) absence or (B) presence of 250 µM ascorbic acid, or 0.1% ethanol as the solvent control, for 40 h, at 20°C. L4-stage worms were subsequently incubated in K-medium with or without phthalates for 24 h at 20°C. Subsequently, intracellular ROS for adult worms were measured using 2′, 7′-dichlorodihydrofluoroscein diacetate. One hundred worms from each exposure condition were used to analyze the intracellular ROS levels. The results are expressed as relative fluorescence units (RFU) of fluorescence relative to 100 worms. The tests were performed a minimum of 3 times. The results were presented as the mean ± standard errors of mean (SEM). Differences compared to the control (0 ppm, 0.1% ethanol) (A) or differences between the populations (B) were considered significant at P<0.05 by one-way ANOVA and the LSD post-hoc test.