GraphPad Statistical Analysis. 

All data on Afternoon of Proestrus 

Time Control 1.56 3.12 

 

1200 0.637 0.07 18 0.44 0.08 5 0.456 0.06 5 

1400 0.906 0.31 13 1.1 0.5 2 0.75 0.18 4 

1600 3.44 0.628 14 1.68 0.175 2 2.94 1.12 4 

1800 21.848 1.75 16 17.53 1.6 11 16.06 1.75 12 

2000 13.445 1.71 18 10.75 3.62 5 8.95 1.25 5 

 

Time 6.25 12.5 25 

1200 0.59 0.06 8 0.717 0.076 11 0.75 0.11 6 

1400 0.942 0.17 12 0.717 0.102 6 1.07 0.25 14 

1600 3.013 1.08 6 2.91 1.3 6 2.94 1.44 11 

1800 12.33 1.6 17 11.88 3.4 7 8.5 2.02 14 

2000 8.67 1.84 8 10 1.65 9 8.39 1.63 13 

 

Time 75 

1200 0.658 0.06 8 

1400 0.891 0.127 6 

1600 4.34 1.19 15 

1800 9.62 1.5 20 

2000 7.7 1.1 17 

Statistical Analysis using GraphPad 

Parameter 

Table Analyzed LH 

 

Two-way ANOVA 

 

Source of Variation % of total variation P value 

 Interaction 5.25 0.0843 

 Column Factor 2.17 0.0283 

 Time 45.16 P<0.0001 

 
Source of Variation P value summary Significant? 

 Interaction ns No 

 Column Factor * Yes 

 Time *** Yes 

 

Source of Variation Df Sum-of-squares Mean square F 

 Interaction 24 803.2 33.47 1.445 

 Column Factor 6 332.4 55.41 2.392 

 Time 4 6905 1726 74.52 

 Residual 313 7251 23.17 

 

Number of missing values -35 

 

Bonferroni posttests 

 

Control vs. 1.56 

Time Control 1.56 Difference 95% CI of diff. 

 1200 0.6370 0.4400 -0.1970 -7.914 to 7.520 

 1400 0.9060 1.100 0.1940 -11.40 to 11.79 

 1600 3.440 1.680 -1.760 -13.30 to 9.779 

 1800 21.85 17.53 -4.318 -10.30 to 1.661 

 2000 13.45 10.75 -2.695 -10.41 to 5.022 

 

Time Difference t P value Summary 

 1200 -0.1970 0.08097 P > 0.05 ns 

 1400 0.1940 0.05307 P > 0.05 ns 

 1600 -1.760 0.4837 P > 0.05 ns 

 1800 -4.318 2.291 P > 0.05 ns 

 2000 -2.695 1.108 P > 0.05 ns 

 

Control vs. 3.12 

Time Control 3.12 Difference 95% CI of diff. 

 1200 0.6370 0.4560 -0.1810 -7.898 to 7.536 

 1400 0.9060 0.7500 -0.1560 -8.884 to 8.572 

 1600 3.440 2.940 -0.5000 -9.154 to 8.154 

 1800 21.85 16.06 -5.788 -11.62 to 0.04129 

 2000 13.45 8.950 -4.495 -12.21 to 3.222 

 

Time Difference t P value Summary 

 1200 -0.1810 0.07439 P > 0.05 ns 

 1400 -0.1560 0.05669 P > 0.05 ns 

 1600 -0.5000 0.1832 P > 0.05 ns 

 1800 -5.788 3.149 P<0.01 ** 

 2000 -4.495 1.847 P > 0.05 ns 

 

Control vs. 6.25 
Time Control 6.25 Difference 95% CI of diff. 

 1200 0.6370 0.5900 -0.04700 -6.533 to 6.439 

 1400 0.9060 0.9420 0.03600 -6.075 to 6.147 

 1600 3.440 3.013 -0.4270 -7.875 to 7.021 

 1800 21.85 12.33 -9.518 -14.83 to -4.201 

 2000 13.45 8.670 -4.775 -11.26 to 1.711 

 

Time Difference t P value Summary 

 1200 -0.04700 0.02298 P > 0.05 ns 

 1400 0.03600 0.01868 P > 0.05 ns 

 1600 -0.4270 0.1818 P > 0.05 ns 

 1800 -9.518 5.677 P<0.001 *** 

 2000 -4.775 2.335 P > 0.05 ns 

 

Control vs. 12.5 

Time Control 12.5 Difference 95% CI of diff. 

 1200 0.6370 0.7170 0.08000 -5.762 to 5.922 

 1400 0.9060 0.7170 -0.1890 -7.723 to 7.345 

 1600 3.440 2.910 -0.5300 -7.978 to 6.918 

 1800 21.85 11.88 -9.968 -16.89 to -3.051 

 2000 13.45 10.00 -3.445 -9.677 to 2.787 

 

Time Difference t P value Summary 

 1200 0.08000 0.04343 P > 0.05 ns 

 1400 -0.1890 0.07956 P > 0.05 ns 

 1600 -0.5300 0.2257 P > 0.05 ns 

 1800 -9.968 4.570 P<0.001 *** 

 2000 -3.445 1.753 P > 0.05 ns 

 

Control vs. 25 

Time Control 25 Difference 95% CI of diff. 

 1200 0.6370 0.7500 0.1130 -7.083 to 7.309 

 1400 0.9060 1.070 0.1640 -5.715 to 6.043 

 1600 3.440 2.940 -0.5000 -6.650 to 5.650 

 1800 21.85 8.500 -13.35 -18.93 to -7.762 

 2000 13.45 8.390 -5.055 -10.61 to 0.5010 

 

Time Difference t P value Summary 

 1200 0.1130 0.04980 P > 0.05 ns 

 1400 0.1640 0.08847 P > 0.05 ns 

 1600 -0.5000 0.2578 P > 0.05 ns 

 1800 -13.35 7.578 P<0.001 *** 

 2000 -5.055 2.886 P < 0.05 * 

 

Control vs. 75 

Time Control 75 Difference 95% CI of diff. 
 1200 0.6370 0.6580 0.02100 -6.465 to 6.507 

 1400 0.9060 0.8910 -0.01500 -7.549 to 7.519 

 1600 3.440 4.340 0.9000 -4.773 to 6.573 

 1800 21.85 9.620 -12.23 -17.35 to -7.108 

 2000 13.45 7.700 -5.745 -10.91 to -0.5825 

 

Time Difference t P value Summary 

 1200 0.02100 0.01027 P > 0.05 ns 

 1400 -0.01500 0.006315 P > 0.05 ns 

 1600 0.9000 0.5032 P > 0.05 ns 

 1800 -12.23 7.575 P<0.001 *** 

 2000 -5.745 3.529 P<0.01 ** 

1200 1400 1600 1800 2000

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Control

25

75

1.56

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6.25

12.5

Time (h)

ng/ml
GRAPHPAD ANALYSIS ON 1800 H DATA ONLY 

1800 h data only 

Time Vehicle 1.56 3.12 

1800 h 21.848 1.75 16 17.53 1.56 11 16.6 1.75 12 

 

Time 6.25 12.5 25 

1800 h 12.33 1.6 17 11.88 3.4 7 8.5 2.02 14 

Time 75 

1800 h 9.6 1.5 20 

 

Parameter Value 

Table Analyzed 

LH 

One-way analysis of variance 

 P value P<0.0001 

 P value summary *** 

 Are means signif. different? (P < 0.05) Yes 

 Number of groups 7 

 F 7.494 

 R squared 0.3332 

 

Bartlett's test for equal variances 

 Bartlett's statistic (corrected) 2.940 

 P value 0.8164 

 P value summary ns 

 Do the variances differ signif. (P < 0.05) No 

 

ANOVA Table SS df MS 

 Treatment (between columns) 2092 6 348.6 

 Residual (within columns) 4186 90 46.52 

 Total 6278 96 

 

Dunnett's Multiple Comparison Test Mean Diff. q P value 95% CI of diff 

 Vehicle vs 1.56 4.318 1.616 P > 0.05 -2.681 to 11.32 

 Vehicle vs 3.12 5.248 2.015 P > 0.05 -1.576 to 12.07 

 Vehicle vs 6.25 9.518 4.007 P < 0.01 3.294 to 15.74 

 Vehicle vs 12.5 9.968 3.225 P < 0.01 1.870 to 18.07 

 Vehicle vs 2513.35 5.348 P < 0.01 6.809 to 19.89 

 Vehicle vs 7512.25 5.354 P < 0.01 6.255 to 18.24 
LH: 1800 hours

Vehicle

1.563.126.2512.5

2575

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ATR dose