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Anti-Fungal Innate Immunity in C. elegans Is Enhanced by Evolutionary Diversification of Antimicrobial Peptides

Figure 3

Induction and regulation of nlp gene expression.

A–C Quantification of the expression of the genes in the nlp-29 locus by qRT-PCR after 24 h of infection by D. coniospora (A), 2 h after needle wounding (B) and 6 h of osmotic stress in liquid (C). + is wild type; – pmk-1(km25). The columns show the average expression level (arbitrary units, +/− SEM) for at least 3 experiments. The level of nlp-27 expression in control animals is set at 1024 (see Materials and Methods). Constitutive expression of the nlp genes can vary across experiment due to differences in the exact age of the worms or conditions (solid or liquid culture). Within a single experiment, age-matched worms were used. D–F Quantification with the Biosort of the normalized fluorescent ratio (green/red) of worms carrying the integrated frIs7 transgene that contains pnlp-29::GFP and pcol-12::DsRed reporters [19] after 6 h in liquid culture in the presence of increasing concentrations of NaCl (D) and following a osmotic stress in 300 mM NaCl (E & F; see Materials and Methods). The fluorescent ratio in different backgrounds, sek-1(ag1), sek-1(km4), nsy-1(ky397), pmk-1(km25) or tir-1(tm3036), for worms after osmotic stress (purple) is compared to control worms (blue). As explained more fully elsewhere [19], due to the nature of the distribution, standard deviations are not an informative parameter and are not shown on this or subsequent figures using the Biosort. The number of worms used in each test is shown in parenthesis. The results shown are representative of at least 3 independent experiments.

Figure 3

doi: https://doi.org/10.1371/journal.ppat.1000105.g003