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. 2020 May 25;11:760. doi: 10.3389/fphar.2020.00760

Figure 5.

Figure 5

Circumdatin D delayed nematode paralysis upon temperature up-shift via suppression AChE and inflammatory-related gene expression in CL4176 transgenic C. elegans strains. (A) Timeline illustrating the time at which the temperature was raised from 16 to 25°C in egg-synchronized CL4176 worms, when worms were fed on circumdatin D-containing NGM medium and when the paralysis assay was scored. (B) The paralysis assays were quantified in the transgenic CL4176 strain treated with or without different concentrations of circumdatin D. Egg-synchronized CL4176 worms were incubated at 16°C for 48 h, treated with appropriate concentrations of circumdatin D at 16 °C for 36 h, then up-shifted to 25°C, and harvested at 36 h after temperature being up-shifted. Data are shown as percentages ± SD of worms paralyzed (n = 90, three independent assays). (C) Effect of circumdatin D on AChE activities, as measured by modified Ellman's spectrophotometric method in CL4176 transgenic C. elegans. Data are normalized with respect to control, and values are the mean ± SD of three independent experiments. (D) Real-time qRT-PCR analysis of AChE and inflammation -associated gene expression in CL4176 transgenic C. elegans. Total RNA was isolated from worms 36 h post temperature up-shift and subjected to DNase digestion. The gene expression levels were normalized to F23B2.13 mRNA. Values represent the mean ± SD of three independent experiments (#compared with blank, *compared with the control, *P < 0.05, **P < 0.01, ***P < 0.001).