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Supplementary Fig. 1. Effect of Kiss2 administration on anxiety-like behaviour and serotonergic genes. A: Side view of behavioural tracking of an adult male zebrafish subjected to either distilled water (control) or Kiss2 (10-11mol/fish, intracranial injection) for 8-min recording 4 hours post administration. B-D: Graphs of the duration of freezing (B), the number of top to bottom transitions (C) and the total distance moved (D) in the adult zebrafish injected with either distilled water (control, open bars) or Kiss2 (closed bars). E-G: Graphs of relative mRNA levels of tph2 (E), pet1 (F) and slc6a4a (G) in the brain of adult zebrafish injected with either distilled water or Kiss1 at 6 hours post administration. Data are presented as mean ± SEM. Independent t-test and the post-hoc Tukey test comparisons between control and Kiss2 injected zebrafish.
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Supplementary Fig. 2. Effect of a single AS exposure on kiss1 and 5-HT related genes. Graphs showing the effect of a single exposure of AS (8-min) on mRNA levels of kiss1 (A), pet1 (B), tph2 (C) slc6a4a (D) genes (n =5/group). Data are presented as mean ± SEM. Independent t-test comparisons between control and AS-exposed fish.
Supplementary Fig. 3. Binding selectivity of Kiss1-SAP to Kiss-R types by luciferase reporter assay. Graphs showing induction firefly luciferase activity divided by Renilla luciferase activity in the HEK 293T cells expressing either Kiss-R1 or Kiss-R2 by Blank-SAP, Kiss1-SAP and zebrafish kisspeptin1-15 (10-5M). Kiss1-SAP and kisspeptin1-15 induced similar luciferase activities in both Kiss-R1 and Kiss-R2 expressing cells. Luciferase activity induced by Kiss1-SAP and kisspeptin1-15 was significantly higher (2.2- and 2.1-fold, P=0.055 and P=0.018) in Kiss-R1 expressing cells as compared to Kiss-R2. Data are presented as mean ± SEM. ANOVA followed by Fisher’s PLSD test among Blank-SAP, Kiss1-SAP and kisspeptin1-15, and independent t-test comparisons for Kiss1-SAP and kisspeptin1-15 between Kiss-R1 and Kiss-R2.
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Submission PDF
Submission PDF
Supplementary Fig. 1. Effect of Kiss2 administration on anxiety-like behaviour and serotonergic genes. A: Side view of behavioural tracking of an adult male zebrafish subjected to either distilled water (control) or Kiss2 (10-11mol/fish, intracranial injection) for 8-min recording 4 hours post administration. B-D: Graphs of the duration of freezing (B), the number of top to bottom transitions (C) and the total distance moved (D) in the adult zebrafish injected with either distilled water (control, open bars) or Kiss2 (closed bars). E-G: Graphs of relative mRNA levels of tph2 (E), pet1 (F) and slc6a4a (G) in the brain of adult zebrafish injected with either distilled water or Kiss1 at 6 hours post administration. Data are presented as mean ± SEM. Independent t-test and the post-hoc Tukey test comparisons between control and Kiss2 injected zebrafish.
Submission PDF
Supplementary Fig. 2. Effect of a single AS exposure on kiss1 and 5-HT related genes. Graphs showing the effect of a single exposure of AS (8-min) on mRNA levels of kiss1 (A), pet1 (B), tph2 (C) slc6a4a (D) genes (n =5/group). Data are presented as mean ± SEM. Independent t-test comparisons between control and AS-exposed fish.
Supplementary Fig. 3. Binding selectivity of Kiss1-SAP to Kiss-R types by luciferase reporter assay. Graphs showing induction firefly luciferase activity divided by Renilla luciferase activity in the HEK 293T cells expressing either Kiss-R1 or Kiss-R2 by Blank-SAP, Kiss1-SAP and zebrafish kisspeptin1-15 (10-5M). Kiss1-SAP and kisspeptin1-15 induced similar luciferase activities in both Kiss-R1 and Kiss-R2 expressing cells. Luciferase activity induced by Kiss1-SAP and kisspeptin1-15 was significantly higher (2.2- and 2.1-fold, P=0.055 and P=0.018) in Kiss-R1 expressing cells as compared to Kiss-R2. Data are presented as mean ± SEM. ANOVA followed by Fisher’s PLSD test among Blank-SAP, Kiss1-SAP and kisspeptin1-15, and independent t-test comparisons for Kiss1-SAP and kisspeptin1-15 between Kiss-R1 and Kiss-R2.
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