Table 1.
Statistical analysis of behavioral experiments
| Figure panel | Assay | Statistical test; findings | Post hoc analysis (adjusted p values) | No. of subjects |
|---|---|---|---|---|
| 5A, B | Neurological examination and tail suspension | χ2 test | NA | Control: n = 12 |
| Observed versus expected (10% failure rate) | cKO: n = 9 | |||
| Control: | ||||
| All tests: p > 0.9999 | ||||
| cKO: | ||||
| All tests: p > 0.9999 | ||||
| 5C | Negative geotaxis | Mann–Whitney U test | NA | Control: n = 12 |
| Control versus cKO | cKO: n = 9 | |||
| p = 0.3100 | ||||
| 5D | Rotarod | Mann–Whitney U test | NA | Control: n = 12 |
| Control versus cKO | cKO: n = 9 | |||
| p = 0.5071 | ||||
| 5E | Wire hang | Mann–Whitney U test | NA | Control: n = 12 |
| Control versus cKO | cKO: n = 9 | |||
| p = 0.1166 | ||||
| 5F | Novel odor recognition | Two-way repeated-measures ANOVA | Wilcoxson matched-pairs signed rank test | Control: n = 18 |
| Odor 1: | Control: | cKO: n = 16 | ||
| Genotype: p = 0.1205 | Trial 3 versus Trial 4: p = 0.0210 | |||
| Trial number: p < 0.0001 | Trial 6 versus Trial 7: | |||
| Interaction: p = 0.0450 | p = 0.0176 | |||
| Odor 2: | cKO: | |||
| Genotype: p = 0.2030 | Trial 3 versus Trial 4: p = 0.0459 | |||
| Trial number: p < 0.0001 | Trial 6 versus Trial 7: | |||
| Interaction: p = 0.9217 | p = 0.0459 | |||
| 6A, 7D | Hot plate | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 24 |
| Genotype: p < 0.0001 | Pretamoxifen versus 7 weeks post-tamoxifen | cKO: n = 18 | ||
| Time: p = 0.2376 | Control: p = 0.0014 | |||
| Interaction: p < 0.0001 | cKO: p < 0.0001 | |||
| 6B, 7E | Tail immersion | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 22 |
| Genotype: p < 0.0001 | Pretamoxifen versus 7 weeks post-tamoxifen | cKO: n = 16 | ||
| Time: p < 0.0001 | Control: p = 0.8191 | |||
| Interaction: p < 0.0001 | cKO: p < 0.0001 | |||
| 6C, 7F | Hargreaves test | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 24 |
| Genotype: p < 0.0001 | Pretamoxifen versus 7 weeks post-tamoxifen | cKO: n = 18 | ||
| Time: p < 0.0001 | Control: p = 0.2694 | |||
| Interaction: p = 0.0004 | cKO: p < 0.0001 | |||
| 6D | Randall-Selitto test | Mann–Whitney U test | NA | Control: n = 12 |
| Control versus cKO | cKO: n = 9 | |||
| p < 0.0001 | ||||
| 6E | Intraplantar capsaicin | Mann–Whitney U test | NA | Control: n = 8 |
| Control versus cKO | cKO: n = 11 | |||
| p < 0.0001 | ||||
| 6F | Acetic acid (i.p.) | Mann–Whitney U test | NA | Control: n = 12 |
| Control versus cKO | cKO: n = 8 | |||
| p = 0.0032 | ||||
| 6G, H | Formalin test, nocifensive behavior | Two-way ANOVA | Sidak's multiple-comparisons test | Control: n = 12 |
| Genotype: p = 0.0061 | Control versus cKO | cKO: n = 9 | ||
| Phase: p < 0.0001 | Phase I: p = 0.0069 | |||
| Interaction: p = 0.1405 | Phase II: p = 0.5503 | |||
| 6I | Formalin test, latency to onset | Mann–Whitney U test | NA | Control: n = 12 |
| Control versus cKO | cKO: n = 9 | |||
| p < 0.0001 | ||||
| 6J | Chloroquine scratching | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | |
| Genotype: p = 0.4825 | Pretamoxifen versus 8 weeks post-tamoxifen | |||
| Treatment: p = 0.0005 | Control: p = 0.5450 | |||
| Interaction: p = 0.0119 | cKO: p = 0.0003 | |||
| 6K | Histamine scratching | Mann–Whitney U test | NA | Control: n = 12 |
| Control versus cKO | cKO: n = 11 | |||
| p = 0.0381 | ||||
| 7A | von Frey threshold | Two-way repeated-measures ANOVA | NA | Control: n = 23 |
| Genotype: p = 0.0554 | cKO: n = 18 | |||
| Time: p = 0.6945 | ||||
| Interaction: p = 0.3343 | ||||
| 7B | Shock intensity-response relationship | Two-way ANOVA | Sidak's multiple-comparisons test | Control: n = 11 |
| Genotype: p = 0.1133 | Control versus cKO | cKO: n = 9 | ||
| Intensity: p < 0.0001 | 0.1 mA: p = 0.4528 | |||
| Interaction: p = 0.8191 | 0.3 mA: p = 0.9998 | |||
| 0.5 mA: p = 0.8528 | ||||
| 0.7 mA: p = 0.9356 | ||||
| 7C | Active avoidance learning | Two-way repeated-measures ANOVA | NA | Control: n = 12 |
| Genotype: p = 0.9687 | cKO: n = 8 | |||
| Block: p < 0.0001 | ||||
| Interaction: p = 0.2074 | ||||
| 8A | Fos induction, heat stimulus, laminae I-II | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 4 |
| Genotype: p = 0.0034 | Ipsilateral versus contralateral | cKO: n = 4 | ||
| Side: p = 0.0037 | Control: p = 0.0017 | |||
| Interaction: p = 0.0064 | cKO: p = 0.9275 | |||
| 8A | Fos induction, heat stimulus, lamina V | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 4 |
| Genotype: p = 0.1145 | Ipsilateral versus contralateral | cKO: n = 4 | ||
| Side: p = 0.0026 | Control: p = 0.0030 | |||
| Interaction: p = 0.0283 | cKO: p = 0.3537 | |||
| 8B | Fos induction, mechanical stimulus, laminae I-II | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 4 |
| Genotype: p = 0.0005 | Ipsilateral versus contralateral | cKO: n = 4 | ||
| Side: p = 0.0002 | Control: p < 0.0001 | |||
| Interaction: p = 0.0003 | cKO: p = 0.9443 | |||
| 8B | Fos induction, mechanical stimulus, lamina V | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 4 |
| Genotype: p = 0.9883 | Ipsilateral versus contralateral | cKO: n = 4 | ||
| Side: p = 0.2833 | Control: p = 0.0059 | |||
| Interaction: p = 0.0013 | cKO: p = 0.0393 | |||
| 8C | Fos induction, 2% formalin (i.pl.), laminae I-II | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 4 |
| Genotype: p = 0.0105 | Ipsilateral versus contralateral | cKO: n = 4 | ||
| Side: p = 0.0058 | Control: p = 0.0109 | |||
| Interaction: p = 0.1199 | cKO: p = 0.2700 | |||
| 8C | Fos induction, 2% formalin (i.pl.), lamina V | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 4 |
| Genotype: p = 0.4297 | Ipsilateral versus contralateral | cKO: n = 4 | ||
| Side: p = 0.0459 | Control: p = 0.2477 | |||
| Interaction: p = 0.9615 | cKO: p = 0.2260 | |||
| 8D | Fos induction, electrical stimulus, laminae I-II | Unpaired t test | NA | Control stimulated: n = 4 |
| Stimulated versus unstimulated subjects | Control unstimulated: n = 3 | |||
| Control: p = 0.0020 | cKO stimulated: n = 5 | |||
| cKO: p = 0.0038 | cKO unstimulated: n = 4 | |||
| 8D | Fos induction, electrical stimulus, lamina V | Unpaired t test | NA | Control stimulated: n = 4 |
| Stimulated versus unstimulated subjects | Control unstimulated: n = 3 | |||
| Control: p = 0.6420 | cKO stimulated: n = 5 | |||
| cKO: p = 0.1221 | cKO unstimulated: n = 4 | |||
| 9A | Acetone test, SNI pretamoxifen | Two-way repeated-measures ANOVA | Unpaired t test with Holm–Sidak multiple-comparisons correction | Control: n = 17 |
| Genotype: p = 0.0038 | Control versus cKO | cKO: n = 17 | ||
| Time: p = 0.0028 | BL: p = 0.9099 | |||
| Interaction: p = 0.0080 | D1: p = 0.9210 | |||
| D3: p = 0.9210 | ||||
| W1: p = 0.9210 | ||||
| W2: p = 0.9210 | ||||
| W4: p = 0.0095 | ||||
| W5: p = 0.0499 | ||||
| W7: p = 0.0499 | ||||
| W8: p = 0.0849 | ||||
| W9: p = 0.1101 | ||||
| W10: p = 0.1101 | ||||
| 9B | von Frey threshold, SNI pretamoxifen | Two-way repeated-measures ANOVA | NA | Control: n = 17 |
| Genotype: p = 0.1403 | cKO: n = 17 | |||
| Time: p < 0.0001 | ||||
| Interaction: p = 0.8480 | ||||
| 9C | Acetone test, SNI post-tamoxifen | Two-way repeated-measures ANOVA | Unpaired t test with Holm–Sidak multiple-comparisons correction | Control: n = 22 |
| Genotype: p = 0.0005 | Control versus cKO | cKO: n = 17 | ||
| Time: p = 0.0009 | BL: p = 0.3505 | |||
| Interaction: p = 0.2335 | W0.5: p = 0.0066 | |||
| W1: p = 0.0016 | ||||
| W2: p = 0.0028 | ||||
| 9D | von Frey threshold, SNI post-tamoxifen | Two-way repeated-measures ANOVA | NA | Control: n = 22 |
| Genotype: p = 0.5078 | cKO: n = 17 | |||
| Time: p < 0.0001 | ||||
| Interaction: p = 0.9924 | ||||
| 9F | Microgliosis, pixel density | Two-way ANOVA | NA | Control: n = 4 |
| Genotype: p = 0.0761 | cKO: n = 4 | |||
| Side: p = 0.0157 | ||||
| Interaction: p = 0.5383 | ||||
| 9G | Hargreaves test, CFA model | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 12 |
| Genotype: p < 0.0001 | Control: | cKO: n = 10 | ||
| Time: p < 0.0001 | BL versus D1: p < 0.0001 | |||
| Interaction: p < 0.0001 | BL versus D2: p < 0.0001 | |||
| BL versus D3: p < 0.0001 | ||||
| BL versus D4: p < 0.0001 | ||||
| BL versus D7: p = 0.0002 | ||||
| cKO: | ||||
| BL versus D1: p > 0.9999 | ||||
| BL versus D2: p = 0.8362 | ||||
| BL versus D3: p = 0.9510 | ||||
| BL versus D4: p = 0.6788 | ||||
| BL versus D7: p = 0.9837 | ||||
| 9H | von Frey threshold, CFA model | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 12 |
| Genotype: p = 0.0002 | Control: | cKO: n = 10 | ||
| Time: p < 0.0001 | BL versus D1: p < 0.0001 | |||
| Interaction: p = 0.1999 | BL versus D2: p < 0.0001 | |||
| BL versus D3: p < 0.0001 | ||||
| BL versus D4: p < 0.0001 | ||||
| BL versus D7: p < 0.0001 | ||||
| cKO: | ||||
| BL versus D1: p < 0.0001 | ||||
| BL versus D2: p < 0.0001 | ||||
| BL versus D3: p < 0.0001 | ||||
| BL versus D4: p < 0.0001 | ||||
| BL versus D7: p < 0.0001 | ||||
| 9I | Paw edema, CFA model | Two-way repeated-measures ANOVA | NA | Control: n = 12 |
| Genotype: p = 0.0530 | cKO: n = 10 | |||
| Time: p < 0.0001 | ||||
| Interaction: p = 0.1033 | ||||
| 9J | Fos induction, non-noxious mechanical stimulus (rotarod walking) in CFA model, laminae I-II | Two-way repeated-measures ANOVA Genotype: p = 0.5855 Side: p = 0.0039 Interaction: p = 0.2020 |
Sidak's multiple-comparisons test Ipsilateral versus contralateral Control: p = 0.0110 cKO: p = 0.1344 |
Control: n = 4 cKO: n = 4 |
| 9K | Hargreaves test, incision model | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 12 |
| Genotype: p < 0.0001 | Control: | cKO: n = 10 | ||
| Time: p = 0.0622 | BL versus 2 h: p = 0.0005 | |||
| Interaction: p = 0.0013 | BL versus D2: p = 0.0158 | |||
| BL versus D7: p > 0.9999 | ||||
| cKO: | ||||
| BL versus 2 h: p = 0.9799 | ||||
| BL versus D2: p > 0.9999 | ||||
| BL versus D7: p = 0.9973 | ||||
| 9L | von Frey threshold, incision model | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Control: n = 12 |
| Genotype: p = 0.5975 | Control: | cKO: n = 11 | ||
| Time: p < 0.0001 | BL versus 2 h: p = 0.0018 | |||
| Interaction: p = 0.9560 | BL versus D2: p = 0.0101 | |||
| BL versus D7: p > 0.9999 | ||||
| cKO: | ||||
| BL versus 2 h: p = 0.0131 | ||||
| BL versus D2: p = 0.1028 | ||||
| BL versus D7: p = 0.9743 | ||||
| 9M | Capsaicin-induced secondary mechanical allodynia | Wilcoxon matched-pairs signed rank test | NA | Control: n = 12 |
| BL versus 30 min | cKO: n = 10 | |||
| Control: p = 0.0210 | ||||
| cKO: p = 0.8457 | ||||
| 11A | Hot plate: GX-201 | Unpaired t test | NA | Vehicle: n = 12 |
| Vehicle versus GX-201 | GX-201: n = 9 | |||
| p = 0.0336 | ||||
| 11B | Hot plate: GX-585 | Unpaired t test | NA | Vehicle: n = 8 |
| Vehicle versus GX-585 | GX-585: n = 15 | |||
| p = 0.0064 | ||||
| 11C | Tail immersion: GX-201 | Unpaired t test | NA | Vehicle: n = 12 |
| Vehicle versus GX-201 | GX-201: n = 11 | |||
| p = 0.0113 | ||||
| 11D | Tail immersion: GX-585 | Unpaired t test | NA | Vehicle: n = 8 |
| Vehicle versus GX-585 | GX-585: n = 15 | |||
| p = 0.0114 | ||||
| 11E | Randall–Selitto test: GX-585 | Unpaired t test | NA | Vehicle: n = 8 |
| Vehicle versus GX-585 | GX-585: n = 15 | |||
| p = 0.0036 | ||||
| 11F | Randall–Selitto test: GX-201 | Unpaired t test | NA | Vehicle: n = 12 |
| Vehicle versus GX-201 | GX-201: n = 11 | |||
| p = 0.8214 | ||||
| 11G | Hargreaves test, CFA model: GX-201 | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Vehicle: n = 12 |
| Treatment: p = 0.0469 | Vehicle: | GX-201: n = 12 | ||
| Time: p < 0.0001 | Pre-CFA versus post-CFA, predose: p = 0.0002 | |||
| Interaction: p = 0.0012 | Pre-CFA versus post-CFA, postdose: p = 0.0002 | |||
| Post-CFA, predose versus post-CFA, postdose: | ||||
| p > 0.9999 | ||||
| GX-201: | ||||
| Pre-CFA versus post-CFA, predose: p < 0.0001 | ||||
| Pre-CFA versus post-CFA, postdose: p > 0.9999 | ||||
| Post-CFA, predose versus post-CFA, postdose: | ||||
| p < 0.0001 | ||||
| 11H | Hargreaves test, CFA model: GX-585 | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Vehicle: n = 6 |
| Treatment: p = 0.1107 | Vehicle: | GX-585: n = 6 | ||
| Time: p = 0.0203 | Pre-CFA versus post-CFA, predose: p = 0.1276 | |||
| Interaction: p = 0.8786 | Pre-CFA versus post-CFA, postdose: p = 0.6355 | |||
| Post-CFA, predose versus post-CFA, postdose: | ||||
| p = 0.5098 | ||||
| GX-585: | ||||
| Pre-CFA versus post-CFA, predose: p = 0.0918 | ||||
| Pre-CFA versus post-CFA, postdose: p = 0.9157 | ||||
| Post-CFA, predose versus post-CFA, postdose: | ||||
| p = 0.1885 | ||||
| 11I | Capsaicin-induced secondary mechanical allodynia:GX-585 | Two-way repeated-measures ANOVA | Sidak's multiple-comparisons test | Vehicle: n = 12 |
| Treatment: p = 0.0395 | Preinjection: | 10 mg/kg: n = 12 | ||
| Time: p < 0.0001 | Vehicle versus 10 mg/kg: p = 0.9993 | 30 mg/kg: n = 11 | ||
| Interaction: p = 0.8148 | Vehicle versus 30 mg/kg: p = 0.9785 | 60 mg/kg: n = 11 | ||
| Vehicle versus 60 mg/kg: p = 0.1026 | ||||
| Postinjection: | ||||
| Vehicle versus 10 mg/kg: p > 0.9999 | ||||
| Vehicle versus 30 mg/kg: p = 0.9284 | ||||
| Vehicle versus 60 mg/kg: p = 0.3197 | ||||
| 11J | Novel odor recognition | Two-way repeated-measures ANOVA | Wilcoxson matched-pairs signed rank test | Vehicle: n = 10 |
| Odor 1: | Vehicle: | GX-585: n = 10 | ||
| Treatment: p = 0.0173 | Trial 3 versus Trial 4: p = 0.1797 | |||
| Trial number: p = 0.0476 | Trial 6 versus Trial 7: p = 0.0625 | |||
| Interaction: p = 0.5307 | GX-585: | |||
| Odor 2: | Trial 3 versus Trial 4: p = 0.5000 | |||
| Treatment: p = 0.0012 | Trial 6 versus Trial 7: p = 0.8125 | |||
| Trial number: p = 0.3027 | ||||
| Interaction: p = 0.3586 |