Mean GEQ values P value Directional fold change   1 dph 16 wk 16wk v 1dph (Age) F v RBC2 (Line) Gene F RBC2 F RBC2 Age Line Inter F RBC2 Combined 1 dph 16 wk Combined ABCC9 0.49 ± 0.16 0.44 ± 0.17 0.42 ± 0.24 0.25 ± 0.11 0.116 0.499 0.312 -1.18 -1.75 -1.41 1.11 1.67 1.31 ACTB 1.15 ± 0.16 1.77 ± 1.23 0.94 ± 0.49 0.83 ± 0.14 0.239 0.612 0.959 -1.23 -2.13 -1.67 -1.54 1.13 -1.25 ACTC1 1.24 ± 0.16 1.40 ± 0.46 0.73 ± 0.15 0.71 ± 0.13 0.001* 0.489 0.635 -1.69 -1.96 -1.85 -1.12 1.03 -1.06 DES 1.17 ± 0.29 0.97 ± 0.40 0.49 ± 0.17 0.57 ± 0.35 0.006* 0.786 0.830 -2.38 -1.69 -2.04 1.21 -1.16 1.08 DMD 0.82 ± 0.07 0.60 ± 0.10 0.40 ± 0.16 0.41 ± 0.15 0.003* 0.396 0.285 -2.04 -1.45 -1.75 1.37 -1.03 1.21 GAPDH 0.89 ± 0.15 1.05 ± 0.50 1.18 ± 0.33 1.37 ± 0.31 0.028* 0.753 0.367 1.33 1.30 1.32 -1.18 -1.16 -1.16 GATA4 1.10 ± 0.13 1.24 ± 0.35 0.89 ± 0.25 0.87 ± 0.35 0.212 0.773 0.873 -1.23 -1.43 -1.33 -1.12 1.03 -1.05 HPRT1 0.61 ± 0.12 0.68 ± 0.32 0.72 ± 0.12 0.76 ± 0.12 0.039* 0.699 0.299 1.18 1.11 1.14 -1.11 -1.05 -1.09 MDH1 0.89 ± 0.08 1.22 ± 0.57 0.74 ± 0.18 0.84 ± 0.08 0.198 0.510 0.527 -1.20 -1.47 -1.33 -1.37 -1.12 -1.25 MYBPC3 1.28 ± 0.07 1.16 ± 0.28 0.75 ± 0.12 0.74 ± 0.18 0.001*‡ 0.136‡ 0.013* -1.72 -1.56 -1.64 1.11 1.01 1.07 MYH6 1.01 ± 0.10 0.97 ± 0.11 0.73 ± 0.13 0.65 ± 0.17 0.009* 0.423 0.974 -1.39 -1.47 -1.45 1.04 1.11 1.07 MYH7 0.95 ± 0.62 4.76 ± 7.38 0.48 ± 0.12 1.37 ± 0.37 0.205 0.081 0.123 -2.00 -3.45 -3.13 -5.00 -2.86 -4.35 SCGD 1.17 ± 0.06 1.03 ± 0.44 0.95 ± 0.25 0.62 ± 0.19 0.068 0.012* 0.832 -1.22 -1.67 -1.41 1.13 1.55 1.29 SCN5A 0.86 ± 0.13 0.77 ± 0.05 0.42 ± 0.17 0.46 ± 0.40 0.003* 0.55 0.573 -2.08 -1.89 -1.96 1.12 1.02 1.09 TPM1 0.78 ± 0.20 1.41 ± 1.08 0.71 ± 0.33 0.74 ± 0.09 0.677 0.872 0.985 -1.09 -1.92 -1.52 -1.82 -1.03 -1.45 TPM4 1.14 ± 0.07 1.29 ± 0.70 0.82 ± 0.25 0.94 ± 0.39 0.332 0.760 0.281 -1.39 -1.39 -1.39 -1.12 -1.14 -1.14 TNNC1 0.83 ± 0.31 0.92 ± 0.56 0.84 ± 0.50 1.18 ± 0.20 0.247 0.662 0.236 1.01 1.28 1.15 -1.11 -1.41 -1.27 TNNI1 1.06 ± 0.14 1.28 ± 0.74 0.12 ± 0.15 0.03 ± 0.01 0.000* 0.062 0.731 -9.09 -50.00 -16.67 -1.22 4.52 -1.11 TNNT2 0.79 ± 0.11 0.76 ± 0.30 0.85 ± 0.11 1.02 ± 0.13 0.014*‡ 0.906‡ 0.031* 1.08 1.33 1.20 1.03 -1.20 -1.09 TTN 1.03 ± 0.38 0.83 ± 0.42 0.54 ± 0.14 0.45 ± 0.06 0.013* 0.733 0.855 -1.92 -1.82 -1.89 1.25 1.19 1.23 VCL 1.13 ± 0.33 1.27 ± 0.28 0.72 ± 0.31 0.45 ± 0.22 0.013* 0.363 0.488 -1.59 -2.86 -2.04 -1.12 1.60 1.08

Table 4: Summary of CEQ multiplex analysis of cardac gene expression. For each gene the mean GEQ values (based on 3 individuals each with 3 experimental replicates) are normalized to PLN. Directional fold changes (calculated by dividing the GEQ values) and P values from 2 way ANOVA analysis of GEQ values are included for all comparisons. Significant P values (< 0.05) are indicated by asterisks and ‡ denotes statistical comparisons with significant interaction.