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Fig. 3 | BMC Pharmacology and Toxicology

Fig. 3

From: A C. elegans model of electronic cigarette use: Physiological effects of e-liquids in nematodes

Fig. 3

Vapor distillate effects on development, fecundity, vitality and induction of the SKN-1 stress response pathway in C. elegans. a A schematic of the system used to vaporize and collect condensates of the distilled vapor. A syringe was used to pull vapor from an activated e-cig device, using brief pulls and burn times to recapitulate what a normal user might experience during vaping. The vapor was condensed into an extract in a flask suspended over liquid nitrogen. Nicotine recovery was confirmed by quantitative HPLC analysis of the pre-vaped (−) and vaped (+) e-liquids, as indicated. Synchronized populations of N2-Bristol strain nematode embryos were placed onto OP50 bacterial seeded NGM agar plates supplemented with vaporized, distilled PG (PG) or e-liquids containing V2 Red (R) flavoring ± nicotine, as indicated. The worms were allowed to mature on these plates. b Body length was assessed every day, with the ~60 h data shown. c Brood size was determined over the entire fertile period. d Thrashing rates were measured using post-hoc analysis of video acquired following movement of the worms from plates to liquid M9 media. Each measurement consists of at least three independent experimental replicates. The number of individual worms analyzed per replicate was from 10–20 for body size and 5 for brood size and thrashing. The negative reference control data is replicated here from Fig. 1 for comparison. Two-way ANOVA with Bonferroni post-hoc analysis was used for statistical analysis. A hashtag (#) represents significant differences (p < 0.05) between the negative control (C) and treatments lacking nicotine, and an asterisk (*) is used to represent significant differences (p < 0.05) between the nicotine control (C+) and treatments containing nicotine. Though not denoted, the statistical analysis also indicated a main effect for nicotine, but no interaction between nicotine and the solvent groups. e-j Representative fluorescent images of transgenic worms expressing the SKN-1 reporter Pgst-4::GFP on plates containing: e) 10 mM paraquat, g)a vapor distillate of PG (PG), h) a vapor distillate of PG + nicotine (N), i) a vapor distillate of V2 Red e-liquid (R), or j) a vapor distillate of R + N, as indicated. The negative control is shown in panel F. The images were acquired and processed using identical settings. Average fold induction is indicated on the upper right corner of each panel. The scale bar is 200 μm

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