Picture from Asif, Muhammad Zaka et al. (2021) MicroPubl Biol
Figure 1. Determining the Median Lethal Dose for 1-Hydroxyphenazine in C. elegans: A) A stock plate of PD1074 worms at varying life stages was washed with M9 and transferred to a 15 mL conical tube. The worms were bleach synchronized and the eggs were resuspended and left to grow in M9 for 24 hours. The L1 arrested worms were transferred to 10 cm plates and left to grow to L4 for 35-42 hours. Approximately 15 L4 worms were transferred onto prepared experimental plates and incubated for 7 hours. Using a fluorescent bead assay, the worms were scored as dead or alive. B) Each replicate consisted of 7 plates, 2 controls and 5 toxin plates of varying concentrations. The applied concentrations of 1-HP were 90 x greater than the effective concentrations, which were corrected by the volume of the agar. C) A gravid adult PD1074 worm without exposure to fluorescent beads. D) A gravid adult worm following 5 min exposure to fluorescent beads. E) Mean normalized death (y-axis) plotted against effective 1-HP concentration (x-axis). Data labeled by the same letter (A, B, or C) are not significantly different, while those labeled by different letters are statistically different with a p-value < 0.05. F) A linear fit curve applied to the mean normalized death vs. effective 1-HP concentration yielded the following equation: y=0.25x+5.23. Using this equation, the effective LD50 concentration is 179 uM.