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. 2018 May 9;13(5):e0197128. doi: 10.1371/journal.pone.0197128

Fig 5. Combination screening assay for the NTC-Sat2 with hygromycin B-HygR.

Fig 5

(A) Schematic drawings showing the design of the DA9-specific expression cassette in two strains. The OTL80 strain has HygR and tomm-20::STOP::gfp inserted into chromosome II. Worms of this strain can grow on hygromycin B-containing NGM plates and they each have a TOMM-20::GFP signal in their DA9 neuron. The OTL90 strain has Sat2 and sam-4::STOP::gfp inserted into chromosome V. Worms of this strain can grow on NTC-containing NGM plates and they each have a SAM-4::GFP signal in their DA9 neuron. (B) Schematic drawing showing the experimental procedure and expected result. T; TOMM-20::GFP, S; SAM-4::GFP, +: wild-type allele. After crossing, the L4 stages of the F1 generation worms were placed onto selection plates containing 0.04 mg/mL of NTC and/or 0.08 mg/mL of hygromycin B. If NTC-Sat2 selection and hygromycin B-HygR selection work independently, the phenotypes of the F2 generation follow the Mendelian inheritance pattern shown. (C) Observed data: graph showing the phenotype ratios of the F2 worms. The numbers of worms counted are shown in the bars. The ratios obtained in this experiment were almost the same as those expected by Mendelian genetics.