Fig. 1: Pld-1 RNAi promotes ROS formation in nematodes and pld-1 expression inversely correlates with aging.

a Effect of pld-1 RNAi on pld-1 expression in nematodes. Lysates from nematodes at day 1 of adulthood fed with pld-1 RNAi or control RNAi were utilized in RT-PCR using specific pld-1 primers. PCR products were resolved on a 1.5% agarose gel. Representative data from three independent experiments are shown. act-2 mRNA served as the loading control. Nematodes fed with pld-1 RNAi or control RNAi were incubated with DCFDA (b) or DHE (c). The fluorescence intensity of nematodes at day 1 or 8 of adulthood (upper panels) was quantified using ImageJ software by determining the average pixel intensity (n > 116 per day 1 condition; n > 73 per day 8 condition). Data are shown as the means ± SEM. *P < 0.05, **P < 0.01. Representative images of nematodes were obtained at 10× magnification (bottom panels). d Age-dependent reduction in pld-1 mRNA levels. Lysates from nematodes at days 1 and 12 of adulthood were utilized in RT-PCR using specific pld-1 primers. PCR products were resolved on a 1.5% agarose gel (upper panel). Representative data from three independent experiments are shown. act-2 mRNA served as the loading control. The graph shows the relative mRNA level of pld-1 (lower panel). Data are shown as the means ± SEM. ***P < 0.001