dgk-1 mutation suppressed the forgetting defect of the tir-1 loss-of-function mutant. A, Suppression of the forgetting defect of the tir-1 mutant by dgk-1 mutation. Chemotaxis of naive, adapted, and 4 h recovered animals was analyzed. Asterisks indicate significant differences between after adaptation and after 4 h recovery. WT: p = 3.35 × 10−7; tir-1;dgk-1(qj203): p = 4.29 × 10−8 (n = 6, F(4,45) = 9.98, p = 7.18 × 10−6). B, Transgenic rescue of tir-1(tm3036);dgk-1(qj203) animals expressing WT dgk-1 genomic fragments. Asterisks indicate significant differences between after adaptation and after 4 h recovery or between recovery with or without dgk-1 fragment. WT: p = 2.84 × 10−17; tir-1;dgk-1(qj203) without dgk-1 fragment: p = 5.23 × 10−10; recovery with or without dgk-1 fragment: p = 4.08 × 10−5 (n = 10, F(6,108) = 22.22, p = 6.72 × 10−17). C, dgk-1(nu62) and D, dgk-1(sy428) mutations also suppressed tir-1's forgetting defect. Asterisks indicate significant differences between after adaptation and after 4 h recovery. C, WT: p = 2.32 × 10−15; tir-1;dgk-1(nu62): p = 9.38 × 10−6 (n ≥ 10, F(4,135) = 17.42, p = 1.50 × 10−11). D, WT: p = 1.53 × 10−15; tir-1;dgk-1(sy428): p = 0.000247 (n = 8, F(4,63) = 22.51, p = 1.40 × 10−11). E, Representative images of the GFP-tagged ODR-10 protein at cilia of AWA in WT and mutants. Scale bar, 10 μm. F, Quantitative analyses of fluorescent intensities of expression of the GFP-tagged ODR-10 protein at cilia. Statistics: one-way ANOVA with Tukey–Kramer multiple test. Asterisks indicate significant differences between dgk-1(qj203) and others. dgk-1(qj203) versus WT: p = 9.14 × 10−13; versus tir-1(tm3036): p = 9.37 × 10−13; versus tir-1;dgk-1: p = 9.52 × 10−13 (n = 72, 77, 64, 134, F(3,343) = 44.42, p = 2.82 × 10−24). G, Phenotype of the dgk-1(qj203) single mutant. Chemotaxis of naive, adapted, and 4 h recovered animals was analyzed. Asterisks indicate significant differences between after adaptation and after 4 h recovery. WT: p = 1.44 × 10−9; dgk-1(qj203): p = 5.09 × 10−9 (n = 8, F(4,63) = 6.67, p = 0.000153). H, dgk-1 also suppresses the forgetting defect of the tir-1 mutant, even, when attracted by an even lower concentration (1:1000) of diacetyl. Asterisks indicate significant difference between naive and after recovery. tir-1(tm3036): p = 8.91 × 10−5 (n = 12, F(6,132) = 4.06, p = 0.000907). I, dgk-1 only modestly, but significantly, suppresses the forgetting defect in tir-1 in the case of the memory of isoamyl alcohol. Asterisks indicate significant differences between after adaptation and after recovery or naive and after recovery. Adaptation versus recovery in WT: p = 2.54 × 10−18; tir-1(tm3036): p = 9.96 × 10−12; dgk-1(qj203): p = 7.76 × 10−26; tir-1;dgk-1: p = 3.03 × 10−5. Naive versus recovery in tir-1: p = 1.17 × 10−27; tir-1;dgk-1: p = 2.11 × 10−9 (n = 12, F(6,132) = 45.34, p = 8.98 × 10−30). J, tir-1 does not show a forgetting defect in the memory of pyrazine. Asterisks indicate significant differences between after adaptation and after recovery. WT: p = 3.73 × 10−14; tir-1(tm3036): p = 3.36 × 10−10; dgk-1(qj203): p = 3.36 × 10−9; tir-1;dgk-1: p = 3.35 × 10−7 (n = 6, F(6,60) = 2.04, p = 0.0744). Box plots: center line indicates median. Box range, 25th-75th percentiles. Whiskers represent minimum-maximum values. Dots represent the data points. External dots from whisker represent outliers. **p < 0.01; ***p < 0.001; two-way ANOVA with Bonferroni correction (except F).
