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. 2018 Jul 4;38(27):6114–6129. doi: 10.1523/JNEUROSCI.1611-17.2018

Figure 3.

Figure 3.

CMK-1 acts in ASE neurons to regulate learning. A, CMK-1 is expressed in ASE sensory neurons. GFP expressed under the flp-6 promoter (known to drive expression in ASE neurons) colocalizes with mCherry expressed under the cmk-1 promoter in both ASE neurons of wild-type worms. Shown are the ASEL (left) and ASER (right) neurons from the same worm imaged using epifluorescence microscopy. Worms are oriented anterior side to the left and dorsal side to top of the image. Scale bar, 20 μm. Similar results were observed in four (of 4) independent transgenic lines. B, Behavior of cmk-1(ok287) (left) and cmk-1(oy21) (right) mutants expressing wild-type CMK-1 under neuron-specific promoters. Each circle represents the results of a single assay and bars indicate mean ± SD; open and closed circles indicate conditioning in the absence and presence of 20 mm NaCl, respectively. Data were pooled from two independent experiments, where each experiment consisted of three to six plates per condition. Left, A two-way ANOVA across all genotypes and conditions revealed a significant interaction between genotype and condition (F(3,220) = 51, p < 0.0001). A two-way ANOVA between cmk-1(ok287) and cmk-1(ok287);flp-6p::cmk-1(WT)::GFP revealed a significant interaction between genotype and condition (F(1,116) = 48.67, p < 0.0001). Similar results were observed in animals carrying a second, independent cmk-1(ok287); flp-6p::cmk-1(WT)::GFP transgene, which had an average chemotaxis index of −0.64 ± 0.12 (mean ± SD, n = 6) and 0.28 ± 0.45 (mean ± SD, n = 6) with and without conditioning in 20 mm NaCl, respectively. A two-way ANOVA between cmk-1(ok287) and cmk-1(ok287);mec-3p::cmk-1(WT)::GFP revealed a nonsignificant interaction between genotype and condition (F(1,110) = 2.318, p = 0.1308). Right, A two-way ANOVA revealed a significant interaction between genotype and condition (F(2,65) = 45.12, p < 0.0001). C, Behavior of cmk-1(ok287) mutants expressing wild-type CMK-1 under ASEL- and ASER-specific promoters. Each circle represents the results of a single assay and bars indicate mean ± SD; open and closed circles indicate conditioning in the absence and presence of 20 mm NaCl, respectively. For clarity, data were pooled from three independent experiments with the following strains: N2, DAG142, DAG712, DAG716, and DAG721, where each experiment consisted of 6 plates per condition. A two-way ANOVA across all genotypes and conditions revealed a significant interaction between genotype and condition (F(4,170) = 29.5, p < 0.0001). Data for independent ASEL, ASER, and ASEL&ASER rescue lines are shown in Figure 3-1. Sidak's posthoc tests were used to assess the effect of conditioning and to compare genotypes in panels B and C. The results are indicated by asterisks: *** and **** denote p < 0.001 and p < 0.0001, respectively and ns denotes not significant.