Experimental conditions and statistics
| Figure | Experimental conditions | Statistical test |
|---|---|---|
| 1B–G | Adult male crickets 1 week after the imaginal molt were isolated for 3 days. Crickets were injected with 20 mM cycloheximide. 1 h later, 5 mM PTX or vehicle were injected to the crickets. 30 min later, brains were dissected for RNA extraction (n = 8 in each group). | Gryllus fra total vehicle vs. Gryllus fra total PTX, U = 23; Gryllus fra-A vehicle vs. Gryllus fra-A PTX, U = 24, Gryllus fra-B vehicle vs. Gryllus fra-B PTX, U = 15; Gryllus jra vehicle vs. Gryllus jra PTX, U = 32; Gryllus egr-B vehicle vs. Gryllus egr-B PTX, U = 11; Gryllus hr38 vehicle vs. Gryllus hr38 PTX, U = 29, Mann-Whitney U test. |
| 2A, B | Adult male crickets 1 week after the imaginal molt were isolated for 3 days. Crickets were injected with (A) 20 mM cycloheximide or (B) saline. 1 h later, 5 mM PTX was injected to the crickets. Brains were dissected for RNA extraction before PTX injection (0 min), or 15, 30, 45, 60, 90, or 120 min after PTX injection (n = 8 in each group). | Effect of pre-treatment: F(1,98) = 0.9604, p = 0.9004; Effect of time: F(6,98) = 9.034, p < 0.0001; interaction: F(6,98) = 0.3634, p = 0.9004, Two-way ANOVA, Dunnett’s post-hoc test. |
| 2C | Same as in Fig. 2A. | H = 29.21, p < 0.0001, Kruskal-Wallis test, Dunn’s post-hoc test. |
| 2D | Adult male crickets 1 week after the imaginal molt were isolated for 3 days. Cricket were fed with 0.5 M sucrose solution 3 times with 5 min intervals. Brains were dissected for RNA extraction before feeding (naïve), or 1 h after feeding (n = 8 in each group). | t = 3.051, df = 8.491, Welch’s unpaired t-test. |
| 2E | Adult male crickets 1 week after the imaginal molt were isolated for 3 days. Weight-matched crickets were introduced into an arena to interact for 5 min, then re-isolated. Brains were dissected for RNA extraction before interaction (naïve), or 1 h after start of interaction (n = 8 in each group). | H = 12.62, p < 0.01, Kruskal-Wallis test, Dunn’s post-hoc test. |
| 5D | Adult male crickets 1 week after the imaginal molt were isolated for 3 days. Then, brains were dissected for RNA extraction (n = 8 in each group). | t = 15.95 df = 7.094, Welch’s unpaired t-test. |
| 5E | Adult male crickets 1 week after the imaginal molt were isolated for 3 days. 5 mM PTX was injected to the crickets. Brains were dissected for RNA extraction before PTX injection (0 min), or 15, 30, 45, 60, 90, or 120 min after PTX injection (n = 8 in each group). | H = 25.87, p < 0.001, Kruskal-Wallis test, Dunn’s post-hoc test. |
| 5F | Same as in Fig. 5C. | H = 39.42, p < 0.0001, Kruskal-Wallis test, Dunn’s post-hoc test. |
| 5G | Pooled expression data of EYFPnls:PEST and Gryllus egr-B in the brain within 120 min after PTX injection (including data presented in Fig. 5C, D), 60 min after vehicle injection, and naïve crickets were analyzed (n = 72, 8, and 8, respectively). | r = 0.8269, p < 0.0001, Pearson’s correlation analysis. |
| 2-1A, E, I | Same as in Fig. 2A. | Gryllus fra-A, F(6,49) = 3.529, p < 0.001; Gryllus fra-B, F(6,49) = 3.327, p < 0.01; Gryllus jra, F(6,49) = 5.305, p < 0.001, One-way ANOVA, Dunnett’s post-hoc test. |
| 2-1B, F, J | Pooled expression data of Gryllus fra-A, fra-B, jra and egr-B in the brain of cycloheximide pretreated crickets within 120 min after PTX injection (including data presented in Figs. 2-1A, E, and I and naïve crickets were analyzed (n = 48 and 8, respectively). | Gryllus fra-A vs. egr-B, r = 0.7856, p < 0.0001; Gryllus fra-B vs. egr-B, r = 0.5250, p < 0.0001; Gryllus jra vs. egr-B, r = 0.6222, p < 0.0001, Pearson’s correlation analysis. |
| 2-1C, G, K | Same as in Fig. 2D. | Gryllus fra-A, U = 13; Gryllus fra-B, U = 7; Gryllus jra, U = 22, Mann-Whitney U test. |
| 2-1D, H, L | Same as in Fig. 2E. | Gryllus fra-A, H = 10.24, p < 0.05; Gryllus fra-B, H = 13.09, p < 0.01; Gryllus jra, H = 2.284, p = 0.5156, Kruskal-Wallis test, Dunn’s post-hoc test. |
| 1-5B, C | Adult male crickets 1 week after the imaginal molt were isolated for 3 days. Crickets were injected with (B) 20 mM cycloheximide or (C) saline. 1 h later, 5 mM PTX was injected to the crickets. Brains were dissected for RNA extraction before PTX injection (0 min), or 15, 30, 45, 60, 90, or 120 min after PTX injection (n = 8 in each group). | Effect of pre-treatment: F(1,98) = 2.060, p = 0.1544; Effect of time: F(6,98) = 14.57, p < 0.0001; interaction: F(6,98) = 0.7991, p = 0.5729, Two-way ANOVA, Šidák’s post-hoc test. |
| 1-5D | Same as in Fig. 2A, B. | Effect of pre-treatment: F(1,98) = 0.5631, p = 0.4548; Effect of time: F(6,98) = 16.11, p < 0.0001; interaction: F(6,98) = 1.733, p = 0.1212, Two-way ANOVA, Šidák’s post-hoc test. |
| 1-5E | Pooled expression data of Gryllus hr38 and Gryllus egr-B in the brain of saline and cycloheximide pretreated crickets within 120 min after PTX injection (including data presented in Figs. 2A, B and Figs. 1-5B, C), and naïve crickets were analyzed (n = 48, 48, and 8, respectively). | Saline pretreated crickets: r = 0.8498, p < 0.0001; cycloheximide pretreated crickets: r = 0.8446; p < 0.0001; all data included: r = 0.8498, p < 0.0001, Pearson’s correlation analysis |
| 1-5F | Same as in Fig. 2D. | t = 2.501, df = 13.18, Welch’s unpaired t-test. |
| 1-5G | Same as in Fig. 2E. | H = 17.61, p < 0.001, Kruskal-Wallis test, Dunn’s post-hoc test. |
| 2-2 | Adult male crickets 1 week after the imaginal molt were isolated for 3 days. Crickets were received injection of PTX, forskolin, TPA, SNAP, anisomycin, A23187, or vehicle. Brains were dissected for RNA extraction 60 min after injection (PTX, n = 8; the other treatments, n = 16). | Gryllus fra-A, H = 62.94, p < 0.0001; Gryllus fra-B, H = 40.22, p < 0.0001; Gryllus jra, H = 56.30, p < 0.0001; Gryllus egr-B, H = 39.41, p < 0.0001; Gryllus hr38, H = 67.07, p < 0.0001, Kruskal-Wallis test, Dunn’s post-hoc test. |