Supplementary Tables

Supplementary Table S1: Normalization factor. Normalization factor (n) was calculated from the geometric mean of three housekeeping genes for each treatment and replicate. Treatments are as follows A (0.5 mg/l Ca2+ non-kairomone); B (1.5 mg/l Ca2+ non-kairomone); C (2.5 mg/l Ca2+ non-kairomone); D (5.0 mg/l Ca2+ non- kairomone); E (0.5 mg/l Ca2+ with kairomone); F (1.5 mg/l Ca2+ with kairomone); G (2.5 mg/l Ca2+ with kairomone); H (5.0 mg/l Ca2+ with kairomone). The numbers beside the treatments denote the replicates. Cycle threshold (CT) is given as an average of the three technical replicates. Housekeeping Gene CT n A1 Gapdh 16.88 21.18 Stx16 23.97 Capon 23.47 A2 Gapdh 17.03 22.81 Stx16 26.65 Capon 26.14 A3 Gapdh 16.30 20.34 Stx16 22.50 Capon 22.92 A4 Gapdh 17.26 21.49 Stx16 23.41 Capon 24.55 B1 Gapdh 16.75 20.62 Stx16 22.59 CAPON 23.15 B2 Gapdh 18.32 22.83 Stx16 25.67 Capon 25.30 B3 Gapdh 16.83 21.12 Stx16 23.16 Capon 24.14 B4 Gapdh 16.59 21.04 Stx16 23.86

1 Capon 23.51 C1 Gapdh 18.08 21.87 Stx16 24.02 Capon 24.07 C2 Gapdh 16.65 20.78 Stx16 23.28 Capon 23.14 C3 Gapdh 17.49 21.27 Stx16 23.22 Capon 23.67 C4 Gapdh 17.25 21.56 Stx16 23.60 Capon 24.60 D1 Gapdh 17.72 22.30 Stx16 24.29 Capon 25.73 D2 Gapdh 17.03 22.24 Stx16 25.50 Capon 25.31 D3 Gapdh 17.19 21.25 Stx16 23.08 Capon 24.18 D4 Gapdh 16.97 21.56 Stx16 23.97 Capon 24.62 E1 Gapdh 17.25 21.28 Stx16 23.68 Capon 23.58 E2 Gapdh 17.34 21.05 Stx16 23.41 Capon 22.97

2 E3 Gapdh 17.04 20.90 Stx16 23.04 Capon 23.25 E4 Gapdh 17.25 22.05 Stx16 25.00 Capon 24.84 F1 Gapdh 17.00 20.84 Stx16 22.86 Capon 23.28 F2 Gapdh 17.41 21.65 Stx16 23.73 Capon 24.55 F3 Gapdh 17.61 22.14 Stx16 25.60 Capon 24.06 F4 Gapdh 18.15 22.77 Stx16 25.95 Capon 25.07 G1 Gapdh 16.57 21.13 Stx16 24.43 Capon 23.31 G2 Gapdh 17.22 21.10 Stx16 23.15 Capon 23.53 G3 Gapdh 16.39 20.50 Stx16 23.10 Capon 22.75 G4 Gapdh 16.97 21.08 Stx16 23.64 Capon 23.33 H1

3 Gapdh 17.05 21.35 Stx16 24.18 Capon 23.58 H2 Gapdh 17.37 21.58 Stx16 24.19 Capon 23.91 H3 Gapdh 17.07 20.72 Stx16 22.61 Capon 23.06 H4 Gapdh 17.50 21.05 Stx16 23.02 Capon 23.14

4 Supplementary Table 2S Fold Change Calculation Data calculations for the treatments and replicates for each target gene. Treatments are as follows A (0.5 mg/l Ca2+ non-kairomone); B (1.5 mg/l Ca2+ non-kairomone); C (2.5 mg/l Ca2+ non-kairomone); D (5.0 mg/l Ca2+ non-kairomone); E (0.5 mg/l Ca2+ with kairomone); F (1.5 mg/l Ca2+ with kairomone); G (2.5 mg/l Ca2+ with kairomone); H (5.0 mg/l Ca2+ with kairomone). The numbers beside the treatments denote the replicates. Treatment D (5.0 Ca2+ -ve Kairomons) used as the calibrator is highlighted. CT stands for Cycle Threshold; n is the normalization factor (geometric mean of three housekeeping genes, see Appendix 2); FC refers to fold change.

ΔΔCT (Δ Δ Ct of CT- Δ CT of 2-ddCT Average Log2 STD SEM Sample CT of Calb n Δ CT calibrator calibrator) (FC) Log2 FC FC Log2 Log 2 Calb A1 22.55 21.18 -1.37 -2.18 -0.81 1.75 0.81 0.58 0.81 0.41 A2 23.89 22.81 -1.08 -2.18 -1.10 2.15 1.10 A3 23.15 20.34 -2.81 -2.18 0.63 0.65 -0.63 A4 22.64 21.49 -1.15 -2.18 -1.03 2.04 1.03 B1 23.49 20.62 -2.88 -2.18 0.70 0.62 -0.70 -0.69 0.44 0.22 B2 26.30 22.83 -3.47 -2.18 1.29 0.41 -1.29 B3 23.79 21.12 -2.67 -2.18 0.49 0.71 -0.49 B4 23.50 21.04 -2.46 -2.18 0.28 0.83 -0.28 C1 24.42 21.87 -2.55 -2.18 0.37 0.78 -0.37 -0.61 0.47 0.24 C2 24.28 20.78 -3.49 -2.18 1.31 0.40 -1.31 C3 23.75 21.27 -2.48 -2.18 0.30 0.81 -0.30 C4 24.22 21.56 -2.66 -2.18 0.48 0.72 -0.48 D1 24.59 22.30 -2.30 -2.18 0.12 0.92 -0.12 0.00 0.35 0.17 D2 23.93 22.24 -1.70 -2.18 -0.48 1.40 0.48 D3 23.77 21.25 -2.52 -2.18 0.33 0.79 -0.33 D4 23.77 21.56 -2.21 -2.18 0.03 0.98 -0.03 E1 23.94 21.28 -2.66 -2.18 0.48 0.72 -0.48 0.54 0.69 0.35 E2 22.16 21.05 -1.10 -2.18 -1.08 2.11 1.08 E3 22.31 20.90 -1.41 -2.18 -0.77 1.71 0.77 E4 23.44 22.05 -1.39 -2.18 -0.79 1.73 0.79 F1 22.93 20.84 -2.09 -2.18 -0.10 1.07 0.10 0.57 0.66 0.33 F2 22.72 21.65 -1.07 -2.18 -1.11 2.16 1.11 F3 23.15 22.14 -1.01 -2.18 -1.17 2.25 1.17 F4 25.05 22.77 -2.28 -2.18 0.10 0.93 -0.10 G1 22.78 21.13 -1.64 -2.18 -0.54 1.45 0.54 0.30 0.57 0.29

5 G2 23.68 21.10 -2.59 -2.18 0.40 0.76 -0.40 G3 21.75 20.50 -1.25 -2.18 -0.93 1.91 0.93 G4 23.14 21.08 -2.06 -2.18 -0.12 1.09 0.12 H1 24.24 21.35 -2.89 -2.18 0.71 0.61 -0.71 -0.82 0.75 0.37 H2 25.05 21.58 -3.46 -2.18 1.28 0.41 -1.28 H3 22.71 20.73 -1.98 -2.18 -0.20 1.15 0.20 H4 24.70 21.05 -3.65 -2.18 1.47 0.36 -1.47 Cuti A1 24.45 21.18 -3.27 -2.24 1.03 0.49 -1.03 -0.78 0.96 0.48 A2 26.05 22.81 -3.24 -2.24 0.99 0.50 -0.99 A3 24.27 20.34 -3.93 -2.24 1.69 0.31 -1.69 A4 23.15 21.49 -1.67 -2.24 -0.57 1.49 0.57 B1 23.99 20.62 -3.38 -2.24 1.14 0.46 -1.14 -0.74 0.97 0.48 B2 24.39 22.83 -1.56 -2.24 -0.68 1.60 0.68 B3 24.83 21.12 -3.71 -2.24 1.47 0.36 -1.47 B4 24.33 21.04 -3.29 -2.24 1.05 0.48 -1.05 C1 27.06 21.87 -5.19 -2.24 2.95 0.13 -2.95 -1.08 1.38 0.69 C2 23.86 20.78 -3.07 -2.24 0.83 0.56 -0.83 C3 24.41 21.27 -3.14 -2.24 0.90 0.54 -0.90 C4 23.44 21.56 -1.88 -2.24 -0.36 1.29 0.36 D1 23.84 22.30 -1.54 -2.24 -0.70 1.63 0.70 0.00 0.58 0.29 D2 24.23 22.24 -1.99 -2.24 -0.25 1.19 0.25 D3 24.04 21.25 -2.78 -2.24 0.54 0.69 -0.54 D4 24.21 21.56 -2.65 -2.24 0.41 0.75 -0.41 E1 25.68 21.28 -4.39 -2.24 2.15 0.22 -2.15 -0.10 1.46 0.73 E2 22.16 21.05 -1.11 -2.24 -1.13 2.19 1.13 E3 23.23 20.90 -2.33 -2.24 0.09 0.94 -0.09 E4 23.61 22.05 -1.55 -2.24 -0.69 1.61 0.69 F1 24.16 20.84 -3.32 -2.24 1.08 0.47 -1.08 -0.25 0.69 0.34 F2 24.40 21.65 -2.75 -2.24 0.51 0.70 -0.51 F3 23.90 22.14 -1.76 -2.24 -0.48 1.39 0.48 F4 24.89 22.77 -2.12 -2.24 -0.12 1.09 0.12 G1 25.05 21.13 -3.92 -2.24 1.68 0.31 -1.68 -1.63 0.87 0.44 G2 26.17 21.10 -5.07 -2.24 2.83 0.14 -2.83 G3 23.90 20.50 -3.40 -2.24 1.16 0.45 -1.16 G4 24.16 21.08 -3.08 -2.24 0.84 0.56 -0.84 6 H1 24.52 21.35 -3.17 -2.24 0.93 0.52 -0.93 -2.27 1.06 0.53 H2 27.21 21.58 -5.62 -2.24 3.38 0.10 -3.38 H3 24.97 20.73 -4.24 -2.24 2.00 0.25 -2.00 H4 26.06 21.05 -5.01 -2.24 2.77 0.15 -2.77 Icp2 A1 22.64 21.18 -1.46 -2.51 -1.06 2.08 1.06 0.46 1.10 0.55 A2 24.61 22.81 -1.80 -2.51 -0.72 1.65 0.72 A3 24.01 20.34 -3.67 -2.51 1.16 0.45 -1.16 A4 22.78 21.49 -1.29 -2.51 -1.22 2.33 1.22 B1 24.88 20.62 -4.27 -2.51 1.75 0.30 -1.75 -2.50 3.61 1.80 B2 24.81 22.83 -1.98 -2.51 -0.53 1.45 0.53 B3 31.34 21.12 -10.23 -2.51 7.71 0.00 -7.71 B4 24.61 21.04 -3.57 -2.51 1.06 0.48 -1.06 C1 25.70 21.87 -3.83 -2.51 1.32 0.40 -1.32 -4.30 2.58 1.29 C2 30.15 20.78 -9.37 -2.51 6.85 0.01 -6.85 C3 26.80 21.27 -5.53 -2.51 3.01 0.12 -3.01 C4 30.09 21.56 -8.53 -2.51 6.01 0.02 -6.01 D1 24.05 22.30 -1.75 -2.51 -0.76 1.70 0.76 0.00 2.26 1.13 D2 22.54 22.24 -0.30 -2.51 -2.21 4.63 2.21 D3 26.91 21.25 -5.66 -2.51 3.14 0.11 -3.14 D4 23.90 21.56 -2.34 -2.51 -0.17 1.13 0.17 E1 24.18 21.28 -2.90 -2.51 0.39 0.77 -0.39 0.08 0.48 0.24 E2 22.86 21.05 -1.81 -2.51 -0.71 1.63 0.71 E3 23.22 20.90 -2.32 -2.51 -0.20 1.15 0.20 E4 24.75 22.05 -2.70 -2.51 0.19 0.88 -0.19 F1 22.98 20.84 -2.14 -2.51 -0.37 1.29 0.37 -1.11 2.93 1.46 F2 24.65 21.65 -3.00 -2.51 0.49 0.71 -0.49 F3 23.59 22.14 -1.45 -2.51 -1.07 2.09 1.07 F4 30.68 22.77 -7.91 -2.51 5.39 0.02 -5.39 G1 23.44 21.13 -2.31 -2.51 -0.21 1.15 0.21 -0.10 0.84 0.42 G2 24.84 21.10 -3.74 -2.51 1.23 0.43 -1.23 G3 22.24 20.50 -1.74 -2.51 -0.77 1.71 0.77 G4 23.74 21.08 -2.66 -2.51 0.15 0.90 -0.15 H1 30.34 21.35 -9.00 -2.51 6.48 0.01 -6.48 -4.90 3.36 1.68 H2 31.34 21.58 -9.76 -2.51 7.25 0.01 -7.25 H3 23.16 20.73 -2.43 -2.51 -0.08 1.06 0.08 7 H4 29.51 21.05 -8.46 -2.51 5.95 0.02 -5.95 Serca A1 19.10 21.18 2.08 2.39 0.31 0.81 -0.31 0.11 0.85 0.43 A2 19.33 22.81 3.49 2.39 -1.10 2.14 1.10 A3 18.77 20.34 1.56 2.39 0.82 0.56 -0.82 A4 18.61 21.49 2.88 2.39 -0.49 1.41 0.49 B1 18.55 20.62 2.07 2.39 0.32 0.80 -0.32 -0.21 0.54 0.27 B2 21.29 22.83 1.55 2.39 0.84 0.56 -0.84 B3 18.85 21.12 2.27 2.39 0.12 0.92 -0.12 B4 18.20 21.04 2.84 2.39 -0.45 1.37 0.45 C1 19.58 21.87 2.29 2.39 0.10 0.93 -0.10 -0.13 0.19 0.10 C2 18.66 20.78 2.12 2.39 0.27 0.83 -0.27 C3 19.17 21.27 2.10 2.39 0.29 0.82 -0.29 C4 19.05 21.56 2.51 2.39 -0.12 1.09 0.12 D1 19.75 22.30 2.55 2.39 -0.16 1.12 0.16 0.00 0.21 0.11 D2 19.95 22.24 2.28 2.39 0.11 0.93 -0.11 D3 19.11 21.25 2.14 2.39 0.25 0.84 -0.25 D4 18.98 21.56 2.58 2.39 -0.19 1.14 0.19 E1 19.52 21.28 1.76 2.39 0.63 0.65 -0.63 -0.06 0.44 0.22 E2 18.38 21.05 2.67 2.39 -0.28 1.22 0.28 E3 18.71 20.90 2.19 2.39 0.20 0.87 -0.20 E4 19.36 22.05 2.69 2.39 -0.30 1.23 0.30 F1 18.45 20.84 2.39 2.39 0.00 1.00 0.00 0.36 0.47 0.23 F2 18.80 21.65 2.85 2.39 -0.46 1.38 0.46 F3 18.78 22.14 3.36 2.39 -0.97 1.96 0.97 F4 20.40 22.77 2.38 2.39 0.01 0.99 -0.01 G1 18.47 21.13 2.66 2.39 -0.28 1.21 0.28 -0.50 0.64 0.32 G2 19.97 21.10 1.13 2.39 1.26 0.42 -1.26 G3 18.79 20.50 1.71 2.39 0.68 0.63 -0.68 G4 19.02 21.08 2.06 2.39 0.33 0.80 -0.33 H1 20.22 21.35 1.12 2.39 1.27 0.42 -1.27 -0.98 0.29 0.14 H2 20.06 21.58 1.52 2.39 0.87 0.55 -0.87 H3 18.97 20.73 1.76 2.39 0.63 0.64 -0.63 H4 19.81 21.05 1.24 2.39 1.15 0.45 -1.15 Ip3R A1 22.76 21.18 -1.58 -2.90 -1.31 2.49 1.31 1.35 0.44 0.22 8 A2 23.79 22.81 -0.98 -2.90 -1.92 3.78 1.92 A3 21.91 20.34 -1.57 -2.90 -1.33 2.51 1.33 A4 23.54 21.49 -2.05 -2.90 -0.85 1.80 0.85 B1 22.17 20.62 -1.55 -2.90 -1.34 2.54 1.34 1.32 0.42 0.21 B2 25.00 22.83 -2.17 -2.90 -0.73 1.66 0.73 B3 22.51 21.12 -1.39 -2.90 -1.50 2.84 1.50 B4 22.25 21.04 -1.21 -2.90 -1.69 3.22 1.69 C1 23.97 21.87 -2.10 -2.90 -0.80 1.74 0.80 0.74 0.07 0.03 C2 23.02 20.78 -2.24 -2.90 -0.65 1.57 0.65 C3 23.44 21.27 -2.17 -2.90 -0.73 1.65 0.73 C4 23.66 21.56 -2.10 -2.90 -0.80 1.74 0.80 D1 26.04 22.30 -3.74 -2.90 0.85 0.56 -0.85 0.00 0.00 0.00 D2 26.40 22.24 -4.16 -2.90 1.27 0.42 -1.27 D3 24.65 21.25 -3.40 -2.90 0.50 0.71 -0.50 D4 21.84 21.56 -0.28 -2.90 -2.61 6.12 2.61 E1 23.73 21.28 -2.44 -2.90 -0.45 1.37 0.45 1.18 0.58 0.29 E2 22.57 21.05 -1.52 -2.90 -1.38 2.60 1.38 E3 22.74 20.90 -1.84 -2.90 -1.06 2.09 1.06 E4 23.12 22.05 -1.07 -2.90 -1.83 3.56 1.83 F1 23.02 20.84 -2.18 -2.90 -0.72 1.65 0.72 1.48 0.76 0.38 F2 23.62 21.65 -1.97 -2.90 -0.93 1.90 0.93 F3 22.99 22.14 -0.85 -2.90 -2.05 4.14 2.05 F4 23.46 22.77 -0.69 -2.90 -2.21 4.63 2.21 G1 22.21 21.13 -1.07 -2.90 -1.82 3.54 1.82 1.03 0.63 0.32 G2 22.76 21.10 -1.66 -2.90 -1.24 2.36 1.24 G3 22.93 20.50 -2.43 -2.90 -0.47 1.38 0.47 G4 23.40 21.08 -2.32 -2.90 -0.57 1.49 0.57 H1 22.76 21.35 -1.42 -2.90 -1.48 2.79 1.48 1.09 0.33 0.17 H2 23.24 21.58 -1.65 -2.90 -1.25 2.37 1.25 H3 22.71 20.73 -1.98 -2.90 -0.92 1.89 0.92 H4 23.21 21.05 -2.16 -2.90 -0.74 1.67 0.74

9 Supplementary Table S3: Raw CT Values. Cycle threshold (CT) values for all the genes, treatments, replicates and technical replicates. Three technical replicates (three qPCR reactions) were conducted per replicate. Treatments are as follows: A (0.5 mg/l Ca2+ non-kairomone); B (1.5 mg/l Ca2+ non- kairomone); C (2.5 mg/l Ca2+ non-kairomone); D (5.0 mg/l Ca2+ non-kairomone); E (0.5 mg/l Ca2+ with kairomone); F (1.5 mg/l Ca2+ with kairomone); G (2.5 mg/l Ca2+ with kairomone); H (5.0 mg/l Ca2+ with kairomone). The numbers beside the treatments denote the replicates. The sample name is written as Gene treatment-replicate. Threshold was set at 0.063416 ΔRn.

Sample (Gene, treatment, replicate) CT 22.286 Calb A1 9 22.527 Calb A1 8 22.845 Calb A1 4 24.438 Calb A2 3 23.753 Calb A2 9 23.478 Calb A2 3 23.222 Calb A3 6 23.177 Calb A3 2 23.044 Calb A3 8 23.081 Calb A4 3 22.360 Calb A4 5 22.480 Calb A4 1 23.516 Calb B1 3 23.655 Calb B1 0 23.308 Calb B1 7 26.426 Calb B2 9

10 26.302 Calb B2 2 26.184 Calb B2 7 23.548 Calb B3 2 23.698 Calb B3 9 24.111 Calb B3 1 23.364 Calb B4 6 23.490 Calb B4 4 23.639 Calb B4 9 24.185 Calb C1 4 24.256 Calb C1 1 24.806 Calb C1 3 24.132 Calb C2 2 24.521 Calb C2 5 24.172 Calb C2 1 23.576 Calb C3 3 23.606 Calb C3 4 24.059 Calb C3 8 24.162 Calb C4 4 24.291 Calb C4 9 24.202 Calb C4 2 25.220 Calb D1 7 24.392 Calb D1 8

11 24.169 Calb D1 6 24.469 Calb D2 1 23.797 Calb D2 5 23.532 Calb D2 9 24.009 Calb D3 7 23.713 Calb D3 9 23.584 Calb D3 9 23.363 Calb D4 5 23.851 Calb D4 2 24.102 Calb D4 8 24.123 Calb E1 7 24.170 Calb E1 4 23.534 Calb E1 2 22.023 Calb E2 6 22.169 Calb E2 5 22.277 Calb E2 5 22.392 Calb E3 6 22.380 Calb E3 4 22.162 Calb E3 4 23.291 Calb E4 4 23.426 Calb E4 3 23.613 Calb E4 2

12 22.949 Calb F1 7 22.769 Calb F1 0 23.056 Calb F1 3 23.207 Calb F2 4 22.440 Calb F2 0 22.505 Calb F2 1 22.967 Calb F3 0 23.152 Calb F3 7 23.338 Calb F3 4 25.059 Calb F4 3 24.849 Calb F4 9 25.248 Calb F4 3 23.220 Calb G1 4 22.763 Calb G1 4 22.348 Calb G1 9 24.096 Calb G2 4 23.609 Calb G2 4 23.344 Calb G2 4 22.044 Calb G3 8 21.713 Calb G3 2 21.495 Calb G3 3 23.011 Calb G4 8

13 23.346 Calb G4 2 23.058 Calb G4 8 24.006 Calb H1 8 24.194 Calb H1 6 24.509 Calb H1 4 24.631 Calb H2 9 25.038 Calb H2 0 25.470 Calb H2 6 22.485 Calb H3 0 22.750 Calb H3 6 22.906 Calb H3 0 25.223 Calb H4 9 24.435 Calb H4 9 24.451 Calb H4 5 23.490 Capon A1 9 23.456 Capon A1 7 23.481 Capon A1 3 27.051 Capon A2 3 26.002 Capon A2 9 25.366 Capon A2 9 23.229 Capon A3 4 22.924 Capon A3 6

14 22.612 Capon A3 8 24.851 Capon A4 2 24.427 Capon A4 9 24.377 Capon A4 0 23.116 Capon B1 0 23.199 Capon B1 6 23.142 Capon B1 9 25.519 Capon B2 0 25.208 Capon B2 7 25.185 Capon B2 6 23.833 Capon B3 8 24.081 Capon B3 1 24.516 Capon B3 5 23.105 Capon B4 5 23.653 Capon B4 4 23.791 Capon B4 5 23.518 Capon C1 7 24.125 Capon C1 0 24.578 Capon C1 1 23.167 Capon C2 7 23.043 Capon C2 6 23.218 Capon C2 5

15 23.185 Capon C3 9 23.499 Capon C3 2 24.348 Capon C3 8 24.889 Capon C4 0 24.806 Capon C4 2 24.108 Capon C4 6 25.854 Capon D1 0 26.013 Capon D1 1 25.348 Capon D1 7 25.855 Capon D2 8 25.053 Capon D2 8 25.027 Capon D2 7 24.276 Capon D3 3 24.096 Capon D3 8 24.194 Capon D3 1 24.309 Capon D4 4 24.572 Capon D4 2 24.981 Capon D4 0 23.970 Capon E1 5 23.226 Capon E1 8 23.571 Capon E1 2 22.897 Capon E2 0

16 22.920 Capon E2 0 23.115 Capon E2 5 23.109 Capon E3 5 23.189 Capon E3 7 23.464 Capon E3 2 24.433 Capon E4 5 24.884 Capon E4 8 25.208 Capon E4 9 23.287 Capon F1 5 23.276 Capon F1 3 23.285 Capon F1 4 25.088 Capon F2 6 24.268 Capon F2 9 24.299 Capon F2 9 23.965 Capon F3 4 24.112 Capon F3 1 24.116 Capon F3 4 24.987 Capon F4 5 25.063 Capon F4 6 25.166 Capon F4 2 23.519 Capon G1 0 23.349 Capon G1 0

17 23.067 Capon G1 4 23.933 Capon G2 4 23.365 Capon G2 3 23.316 Capon G2 8 23.120 Capon G3 0 22.429 Capon G3 3 22.721 Capon G3 7 23.150 Capon G4 4 23.045 Capon G4 0 23.805 Capon G4 5 23.307 Capon H1 8 23.386 Capon H1 2 24.051 Capon H1 6 23.329 Capon H2 5 24.011 Capon H2 9 24.411 Capon H2 1 23.112 Capon H3 6 23.207 Capon H3 5 22.883 Capon H3 5 23.925 Capon H4 4 22.903 Capon H4 8 22.615 Capon H4 2

18 24.500 Cuti A1 3 24.445 Cuti A1 4 24.397 Cuti A1 7 26.451 Cuti A2 3 26.043 Cuti A2 3 25.648 Cuti A2 0 24.429 Cuti A3 0 24.131 Cuti A3 0 24.246 Cuti A3 4 23.518 Cuti A4 7 23.013 Cuti A4 9 22.930 Cuti A4 2 23.804 Cuti B1 8 23.966 Cuti B1 4 24.210 Cuti B1 6 24.691 Cuti B2 8 23.779 Cuti B2 5 24.703 Cuti B2 6 24.627 Cuti B3 4 24.651 Cuti B3 3 25.204 Cuti B3 3 24.135 Cuti B4 2

19 24.138 Cuti B4 0 24.729 Cuti B4 5 26.809 Cuti C1 6 26.769 Cuti C1 8 27.608 Cuti C1 7 23.933 Cuti C2 3 23.823 Cuti C2 9 23.812 Cuti C2 3 24.069 Cuti C3 0 24.395 Cuti C3 2 24.760 Cuti C3 6 23.521 Cuti C4 3 23.349 Cuti C4 4 23.436 Cuti C4 5 24.137 Cuti D1 8 23.821 Cuti D1 8 23.552 Cuti D1 5 24.837 Cuti D2 8 23.977 Cuti D2 1 23.872 Cuti D2 3 24.026 Cuti D3 1 24.239 Cuti D3 0

20 23.843 Cuti D3 8 23.888 Cuti D4 5 24.120 Cuti D4 5 24.607 Cuti D4 4 26.132 Cuti E1 3 25.396 Cuti E1 0 25.501 Cuti E1 4 22.020 Cuti E2 0 22.095 Cuti E2 2 22.364 Cuti E2 3 23.350 Cuti E3 9 23.142 Cuti E3 6 23.197 Cuti E3 6 23.512 Cuti E4 8 23.656 Cuti E4 3 23.646 Cuti E4 8 24.031 Cuti F1 7 24.039 Cuti F1 7 24.404 Cuti F1 4 24.824 Cuti F2 5 24.096 Cuti F2 5 24.277 Cuti F2 1

21 23.753 Cuti F3 5 23.778 Cuti F3 1 24.176 Cuti F3 8 24.809 Cuti F4 4 24.874 Cuti F4 4 24.995 Cuti F4 7 25.336 Cuti G1 5 25.164 Cuti G1 0 24.659 Cuti G1 8 26.340 Cuti G2 7 26.290 Cuti G2 4 25.867 Cuti G2 7 24.080 Cuti G3 9 24.030 Cuti G3 6 23.586 Cuti G3 4 23.953 Cuti G4 3 24.056 Cuti G4 2 24.465 Cuti G4 1 24.333 Cuti H1 8 24.226 Cuti H1 9 24.997 Cuti H1 7 27.279 Cuti H2 9

22 27.214 Cuti H2 7 27.126 Cuti H2 0 25.170 Cuti H3 3 24.748 Cuti H3 8 24.992 Cuti H3 9 26.328 Cuti H4 5 26.038 Cuti H4 3 25.811 Cuti H4 3 16.782 Gapdh A1 5 16.863 Gapdh A1 9 17.009 Gapdh A1 7 17.396 Gapdh A2 4 16.783 Gapdh A2 2 16.934 Gapdh A2 2 16.468 Gapdh A3 8 16.211 Gapdh A3 4 16.238 Gapdh A3 6 17.423 Gapdh A4 9 17.216 Gapdh A4 1 17.146 Gapdh A4 8 16.676 Gapdh B1 7 16.742 Gapdh B1 4

23 16.845 Gapdh B1 5 18.441 Gapdh B2 5 18.229 Gapdh B2 3 18.293 Gapdh B2 4 16.482 Gapdh B3 6 16.997 Gapdh B3 9 17.029 Gapdh B3 0 16.584 Gapdh B4 1 16.487 Gapdh B4 7 16.726 Gapdh B4 4 17.877 Gapdh C1 0 17.996 Gapdh C1 5 18.373 Gapdh C1 0 16.586 Gapdh C2 7 16.610 Gapdh C2 3 16.778 Gapdh C2 8 17.060 Gapdh C3 6 17.574 Gapdh C3 6 17.853 Gapdh C3 1 17.472 Gapdh C4 4 17.357 Gapdh C4 2 16.947 Gapdh C4 9

24 18.135 Gapdh D1 7 17.496 Gapdh D1 5 17.543 Gapdh D1 6 17.398 Gapdh D2 2 16.856 Gapdh D2 6 16.841 Gapdh D2 2 17.092 Gapdh D3 7 17.554 Gapdh D3 4 16.926 Gapdh D3 6 16.647 Gapdh D4 0 17.007 Gapdh D4 9 17.258 Gapdh D4 1 17.640 Gapdh E1 8 17.166 Gapdh E1 3 16.957 Gapdh E1 3 17.335 Gapdh E2 5 17.237 Gapdh E2 3 17.465 Gapdh E2 0 17.125 Gapdh E3 2 17.008 Gapdh E3 6 17.004 Gapdh E3 9 17.205 Gapdh E4 2

25 17.283 Gapdh E4 7 17.289 Gapdh E4 1 16.883 Gapdh F1 6 16.966 Gapdh F1 8 17.151 Gapdh F1 7 17.714 Gapdh F2 2 17.441 Gapdh F2 5 17.075 Gapdh F2 3 16.901 Gapdh F3 6 16.857 Gapdh F3 8 19.071 Gapdh F3 3 18.186 Gapdh F4 5 18.040 Gapdh F4 6 18.226 Gapdh F4 7 16.821 Gapdh G1 9 16.394 Gapdh G1 9 16.501 Gapdh G1 7 17.547 Gapdh G2 6 17.079 Gapdh G2 0 17.056 Gapdh G2 7 16.340 Gapdh G3 0 16.421 Gapdh G3 6

26 16.422 Gapdh G3 4 16.762 Gapdh G4 9 16.946 Gapdh G4 5 17.206 Gapdh G4 9 16.705 Gapdh H1 2 17.052 Gapdh H1 2 17.412 Gapdh H1 6 17.182 Gapdh H2 2 17.162 Gapdh H2 2 17.779 Gapdh H2 6 17.238 Gapdh H3 2 17.015 Gapdh H3 5 16.968 Gapdh H3 6 17.561 Gapdh H4 8 17.446 Gapdh H4 3 22.555 Icp2 A1 1 22.768 Icp2 A1 5 22.595 Icp2 A1 0 25.451 Icp2 A2 6 24.365 Icp2 A2 8 24.006 Icp2 A2 2 24.132 Icp2 A3 2

27 24.283 Icp2 A3 3 23.611 Icp2 A3 7 23.317 Icp2 A4 6 22.520 Icp2 A4 3 22.506 Icp2 A4 5 24.680 Icp2 B1 6 24.905 Icp2 B1 2 25.062 Icp2 B1 0 24.826 Icp2 B2 1 24.676 Icp2 B2 7 24.939 Icp2 B2 3 31.907 Icp2 B3 3 30.303 Icp2 B3 0 31.822 Icp2 B3 1 24.389 Icp2 B4 4 25.021 Icp2 B4 2 24.425 Icp2 B4 9 25.430 Icp2 C1 3 25.525 Icp2 C1 8 26.135 Icp2 C1 6 29.689 Icp2 C2 3 30.580 Icp2 C2 8

28 30.181 Icp2 C2 3 26.229 Icp2 C3 4 26.564 Icp2 C3 3 27.596 Icp2 C3 0 30.078 Icp2 C4 1 30.093 Icp2 C4 3 24.414 Icp2 D1 6 24.016 Icp2 D1 1 23.721 Icp2 D1 1 23.180 Icp2 D2 3 22.436 Icp2 D2 2 22.001 Icp2 D2 9 26.771 Icp2 D3 9 26.950 Icp2 D3 1 27.008 Icp2 D3 8 23.536 Icp2 D4 5 23.902 Icp2 D4 8 24.266 Icp2 D4 7 24.768 Icp2 E1 1 23.762 Icp2 E1 3 24.014 Icp2 E1 2 22.878 Icp2 E2 5

29 22.926 Icp2 E2 6 22.772 Icp2 E2 6 23.416 Icp2 E3 3 23.183 Icp2 E3 7 23.065 Icp2 E3 0 24.376 Icp2 E4 4 24.841 Icp2 E4 0 25.045 Icp2 E4 0 22.620 Icp2 F1 0 23.042 Icp2 F1 0 23.285 Icp2 F1 8 25.214 Icp2 F2 0 24.202 Icp2 F2 2 24.543 Icp2 F2 1 23.196 Icp2 F3 4 23.467 Icp2 F3 0 24.098 Icp2 F3 4 31.015 Icp2 F4 9 29.735 Icp2 F4 6 31.298 Icp2 F4 7 23.010 Icp2 G1 1 23.291 Icp2 G1 4

30 24.024 Icp2 G1 8 25.163 Icp2 G2 4 25.052 Icp2 G2 1 24.310 Icp2 G2 5 22.329 Icp2 G3 2 22.155 Icp2 G3 2 22.247 Icp2 G3 9 23.235 Icp2 G4 7 23.597 Icp2 G4 7 24.387 Icp2 G4 7 30.114 Icp2 H1 7 30.393 Icp2 H1 5 30.524 Icp2 H1 7 31.021 Icp2 H2 1 31.544 Icp2 H2 5 31.466 Icp2 H2 9 23.322 Icp2 H3 3 23.185 Icp2 H3 8 22.978 Icp2 H3 4 29.437 Icp2 H4 1 29.590 Icp2 H4 8 22.755 Ip3R A1 7

31 22.892 Ip3R A1 3 22.645 Ip3R A1 7 24.189 Ip3R A2 3 23.643 Ip3R A2 8 23.537 Ip3R A2 4 22.105 Ip3R A3 7 21.915 Ip3R A3 6 21.700 Ip3R A3 8 23.760 Ip3R A4 8 23.566 Ip3R A4 5 23.287 Ip3R A4 7 22.066 Ip3R B1 6 22.142 Ip3R B1 9 22.299 Ip3R B1 3 25.024 Ip3R B2 7 25.535 Ip3R B2 9 24.443 Ip3R B2 9 22.560 Ip3R B3 5 22.161 Ip3R B3 2 22.807 Ip3R B3 7 22.264 Ip3R B4 8 22.143 Ip3R B4 5

32 22.352 Ip3R B4 1 24.019 Ip3R C1 7 Ip3R C1 23.617 24.263 Ip3R C1 9 Ip3R C2 22.701 23.112 Ip3R C2 1 23.257 Ip3R C2 8 23.323 Ip3R C3 1 23.325 Ip3R C3 4 23.673 Ip3R C3 3 23.722 Ip3R C4 8 23.690 Ip3R C4 5 23.556 Ip3R C4 1 26.392 Ip3R D1 5 26.042 Ip3R D1 6 25.689 Ip3R D1 8 27.021 Ip3R D2 5 26.388 Ip3R D2 2 25.790 Ip3R D2 1 25.014 Ip3R D3 6 24.461 Ip3R D3 1 24.471 Ip3R D3 6 23.875 Ip3R D4 3

33 23.884 Ip3R D4 9 24.289 Ip3R D4 6 24.143 Ip3R E1 6 23.680 Ip3R E1 0 23.356 Ip3R E1 0 22.355 Ip3R E2 0 22.537 Ip3R E2 0 22.816 Ip3R E2 0 23.028 Ip3R E3 3 22.398 Ip3R E3 7 22.791 Ip3R E3 9 23.126 Ip3R E4 1 23.101 Ip3R E4 3 23.120 Ip3R E4 5 23.036 Ip3R F1 2 23.000 Ip3R F1 7 23.014 Ip3R F1 1 24.119 Ip3R F2 4 23.410 Ip3R F2 8 23.327 Ip3R F2 6 22.545 Ip3R F3 8 22.850 Ip3R F3 0

34 23.565 Ip3R F3 5 23.508 Ip3R F4 5 23.574 Ip3R F4 5 23.300 Ip3R F4 5 22.374 Ip3R G1 0 22.266 Ip3R G1 7 21.983 Ip3R G1 7 23.183 Ip3R G2 4 22.541 Ip3R G2 5 22.545 Ip3R G2 5 23.022 Ip3R G3 1 23.086 Ip3R G3 8 22.683 Ip3R G3 3 23.113 Ip3R G4 1 23.390 Ip3R G4 2 23.691 Ip3R G4 7 22.487 Ip3R H1 9 23.011 Ip3R H1 7 22.790 Ip3R H1 6 23.029 Ip3R H2 9 23.230 Ip3R H2 2 23.445 Ip3R H2 3

35 22.832 Ip3R H3 6 22.711 Ip3R H3 6 22.585 Ip3R H3 5 23.707 Ip3R H4 3 23.159 Ip3R H4 6 22.761 Ip3R H4 5 18.652 Serca A1 3 19.212 Serca A1 6 19.448 Serca A1 3 19.853 Serca A2 8 19.181 Serca A2 2 18.940 Serca A2 7 18.929 Serca A3 4 18.800 Serca A3 6 18.590 Serca A3 7 19.028 Serca A4 7 18.528 Serca A4 3 18.264 Serca A4 7 18.458 Serca B1 9 18.547 Serca B1 9 18.631 Serca B1 1 21.558 Serca B2 2

36 21.363 Serca B2 0 20.939 Serca B2 7 18.829 Serca B3 1 18.591 Serca B3 7 19.116 Serca B3 5 18.065 Serca B4 9 18.227 Serca B4 5 18.305 Serca B4 5 19.298 Serca C1 9 19.690 Serca C1 0 19.748 Serca C1 6 18.449 SercaC2 6 18.641 SercaC2 6 18.884 SercaC2 2 18.868 Serca C3 0 19.146 Serca C3 5 19.510 Serca C3 1 19.277 Serca C4 6 19.084 Serca C4 0 18.776 Serca C4 8 20.291 Serca D1 6 19.482 Serca D1 7

37 19.462 Serca D1 3 20.069 Serca D2 7 20.019 Serca D2 8 19.772 Serca D2 0 19.380 Serca D3 3 19.289 Serca D3 5 18.661 Serca D3 8 18.772 Serca D4 6 18.996 Serca D4 3 19.170 Serca D4 3 19.793 Serca E1 7 19.211 Serca E1 9 19.549 Serca E1 5 18.244 Serca E2 4 18.320 Serca E2 2 18.580 Serca E2 2 18.963 Serca E3 4 18.625 Serca E3 7 18.547 Serca E3 8 19.279 Serca E4 4 19.308 Serca E4 7 19.499 Serca E4 5

38 18.443 Serca F1 8 18.483 Serca F1 7 18.419 Serca F1 5 19.121 Serca F2 6 18.515 Serca F2 7 18.755 Serca F2 1 18.544 Serca F3 7 18.783 Serca F3 5 19.009 Serca F3 9 20.263 Serca F4 6 20.270 Serca F4 6 20.663 Serca F4 5 18.377 Serca G1 4 18.535 Serca G1 2 18.496 Serca G1 3 20.246 Serca G2 9 19.936 Serca G2 9 19.722 Serca G2 6 18.968 Serca G3 2 18.873 Serca G3 2 18.531 Serca G3 8 18.802 Serca G4 6

39 19.063 Serca G4 7 19.188 Serca G4 2 19.952 Serca H1 2 20.407 Serca H1 0 20.312 Serca H1 6 19.805 Serca H2 0 20.039 Serca H2 4 20.340 Serca H2 2 19.164 Serca H3 2 19.048 Serca H3 9 18.709 Serca H3 3 20.106 Serca H4 6 19.888 Serca H4 1 19.434 Serca H4 9 23.750 Stx16 A1 4 24.001 Stx16 A1 1 24.172 Stx16 A1 7 27.688 Stx16 A2 2 26.477 Stx16 A2 0 25.800 Stx16 A2 7 22.279 Stx16 A3 0 22.555 Stx16 A3 2

40 22.682 Stx16 A3 7 24.017 Stx16 A4 9 23.352 Stx16 A4 2 22.860 Stx16 A4 3 22.470 Stx16 B1 6 22.497 Stx16 B1 4 22.806 Stx16 B1 5 25.514 Stx16 B2 7 25.840 Stx16 B2 5 22.773 Stx16 B3 3 23.020 Stx16 B3 7 23.710 Stx16 B3 6 23.626 Stx16 B4 9 23.767 Stx16 B4 9 24.204 Stx16 B4 4 23.485 Stx16 C1 9 24.084 Stx16 C1 3 24.498 Stx16 C1 4 23.142 Stx16 C2 8 23.177 Stx16 C2 2 23.520 Stx16 C2 0 23.128 Stx16 C3 7

41 23.328 Stx16 C3 5 23.507 Stx16 C4 6 23.820 Stx16 C4 0 23.475 Stx16 C4 0 24.700 Stx16 D1 6 24.157 Stx16 D1 2 24.041 Stx16 D1 9 26.163 Stx16 D2 3 25.311 Stx16 D2 7 25.030 Stx16 D2 4 23.136 Stx16 D3 6 23.049 Stx16 D3 3 23.075 Stx16 D3 2 23.340 Stx16 D4 0 24.021 Stx16 D4 7 24.571 Stx16 D4 4 24.336 Stx16 E1 5 23.536 Stx16 E1 3 23.170 Stx16 E1 7 23.496 Stx16 E2 9 23.212 Stx16 E2 2 23.527 Stx16 E2 4

42 23.090 Stx16 E3 4 22.930 Stx16 E3 9 23.109 Stx16 E3 3 24.872 Stx16 E4 8 25.073 Stx16 E4 9 25.069 Stx16 E4 0 22.801 Stx16 F1 5 22.778 Stx16 F1 5 23.009 Stx16 F1 7 24.399 Stx16 F2 1 23.673 Stx16 F2 7 23.143 Stx16 F2 8 24.600 Stx16 F3 4 25.592 Stx16 F3 4 26.631 Stx16 F3 9 25.868 Stx16 F4 0 26.016 Stx16 F4 0 25.983 Stx16 F4 9 24.963 Stx16 G1 5 24.179 Stx16 G1 7 24.158 Stx16 G1 8 23.702 Stx16 G2 4

43 22.737 Stx16 G2 5 23.033 Stx16 G2 5 23.115 Stx16 G3 3 23.094 Stx16 G3 3 23.098 Stx16 G3 1 23.132 Stx16 G4 7 23.483 Stx16 G4 5 24.310 Stx16 G4 6 24.128 Stx16 H1 1 24.232 Stx16 H1 4 23.430 Stx16 H2 7 24.067 Stx16 H2 6 25.097 Stx16 H2 3 22.698 Stx16 H3 6 22.663 Stx16 H3 9 22.488 Stx16 H3 6 23.386 Stx16 H4 6 22.878 Stx16 H4 2 22.806 Stx16 H4 4

44 Supplementary Table S4: One-way ANOVA

One-way ANOVA testing if calcium had a significant effect on the gene expression in the presence or absence of kairomones. Changes in gene expression across the calcium gradient were only observed in animals that were simultaneously exposed to (+) kairomones. Significant values are in bold (P < 0.05).

Gene Kairomone presence (+) Pr (>F) and absence (-) Cutie + 0.034 - 0.504 Icp2 + 0.031 - 0.069 Ip3R + 0.728 - 0.196 Serca + 0.010 - 0.829 Calb + 0.004 - 0.659

45 Supplementary Table S5: Tukey multiple comparison test.

Tukey test was conducted on significant one way ANOVA results (table S1) to investigate which specific calcium treatments significantly affected gene expression. Since calcium only effected gene expression when kairomones were present, Tukey test was performed only on kairomone treatments across the calcium gradient. Bold values indicate significant difference in expression between any two calcium treatments (P < 0.05).

Gene Kairomone Compared Ca2+ P presence (+) treatments (mg/L) and absence (-) Cutie + 0.5-1.5 0.997 0.5-2.5 0.229 0.5-5.0 0.056 1.5-2.5 0.300 1.5-5.0 0.078 2.5-5.0 0.823 Icp2 + 0.5-1.5 0.879 0.5-2.5 0.999 0.5-5.0 0.040 1.5-2.5 0.921 1.5-5.0 0.141 2.5-5.0 0.049 Serca + 0.5-1.5 0.617 0.5-2.5 0.580 0.5-5.0 0.075 1.5-2.5 0.104 1.5-5.0 0.008 2.5-5.0 0.507 Calb + 0.5-1.5 0.999 0.5-2.5 0.954 0.5-5.0 0.061 1.5-2.5 0.937 1.5-5.0 0.055 2.5-5.0 0.143

46 47 Supplementary Table S6: T-test

T-test comparison of means between kairomone and non-kairomone treatments at each calcium concentration for the expression of Calb, Cuti, Icp2, Ip3R, and Serca. Bold values (P < 0.05) indicating significant effect of kairomones on gene expression are in bold. Gene Ca2+ (mg/L) P-value Cutie 0.5 0.426 1.5 0.520 2.5 0.495 5.0 0.019 Icp2 0.5 0.611 1.5 0.654 2.5 0.025 5.0 0.178 Ip3R 0.5 0.701 1.5 0.598 2.5 0.437 5.0 0.361 Serca 0.5 0.602 1.5 0.258 2.5 0.287 5.0 0.029 Calb 0.5 0.949 1.5 0.082 2.5 0.006 5.0 0.209