Supplementary Table 1
Average peak KCl responses of DRG neurons responding to constellation pharmacology triggers 10 µM R9 CBD3 0.002% DMSO A6K Trigger(s) average SEM count average SEM count responder Ach 0.553658 0.021938 300 0.619269 0.025057 325 responder AITC 0.637831 0.119105 21 0.83787 0.116019 16 responder ATP 0.787866 0.040487 207 0.81266 0.047168 190 responder Histamine 0.709658 0.079638 36 0.750603 0.079342 35 responder Menthol 0.48204 0.05045 64 0.70498 0.088 48 responder Capsaicin 0.619908 0.027566 393 0.574082 0.02718 432 responder kcl 0.603849 0.013519 1184 0.591719 0.014646 1301 KCL only 0.575246 0.0202 410 0.526446 0.021103 529 all triggers 0 0
ACh 0.554977 0.023589 214 0.634514 0.032048 211 AITC 0.514096 0.154366 4 0.87365 0.188179 4 ATP 0.822789 0.066823 83 0.894669 0.089754 63 Hist 0.445953 0.118529 4 0.540678 0.102158 5 Ment 0.332511 0.029819 38 0.689671 0.127966 23 Cap 0.589266 0.032695 246 0.518077 0.032972 255 ACh/ AITC 0.041186 0 1 0 ACh/ ATP 0.641581 0.08765 25 0.588704 0.084367 19 ACh/ Hist 0.637404 0.224389 5 0.796731 0.125491 9 ACh/ Ment 0.314589 0.049202 4 0.813618 0 1 ACh/ Cap 0.517266 0.078392 28 0.483926 0.060716 51 AITC/ ATP 0 0.449344 0 1
AITC/ Hist 0.532387 0.128886 2 0 AITC/ Ment 0 0.186509 0 1
AITC/ Cap 0 0.478392 0.042441 2
ATP/ Hist 1.278958 0 1 0.512734 0.144363 2 ATP/ Ment 0.635429 0.120964 5 0.327995 0 1 ATP/ Cap 0.855115 0.075348 64 0.853068 0.082822 73 Hist/ Ment 0.515909 0.174641 4 1.455467 0.231283 3 Hist/ Cap 0.656417 0.216999 7 0.389859 0.133844 3 Ment/ Cap 1.159204 0.312253 5 0.706628 0.2144 8 ACh/ AITC/ ATP 0.18241 0 1 0.786368 0 1 ACh/ AITC/ Hist 1.28866 0 1 0 ACh/ AITC/ Ment 0 0
ACh/ AITC/ Cap 0.02651 0 1 0 ACh/ ATP/ Hist 0 0.809036 0 1
ACh/ ATP/ Ment 0 0
ACh/ ATP/ Cap 0.573797 0.203213 8 0.617002 0.070834 17 ACh/ Hist/ Ment 0 0.538063 0.210029 3
ACh/ Hist/ Cap 0.317027 0.182676 3 0.171103 0 1 ACh/ Ment/ Cap 0.205817 0.084275 2 0.371297 0.228513 2 AITC/ ATP/ Hist 0 0
AITC/ ATP/ Ment 0 0
AITC/ ATP/ Cap 0.69392 0.260132 6 0 AITC/ Hist/ Ment 0 0
AITC/ Hist/ Cap 0 0
AITC/ Ment/ Cap 0.642336 0 1 0 ATP/ Hist/ Ment 1.049628 0.147808 2 0.390997 0 1 ATP/ Hist/ Cap 0.588087 0 1 0 ATP/ Ment/ Cap 0.161867 0 1 0 Hist/ Ment/ Cap 0 0.845867 0 1
ACh/ AITC/ ATP/ Hist 0 0.718172 0 1
ACh/ AITC/ ATP/ Ment 0 1.145044 0 1
ACh/ AITC/ ATP/ Cap 0 1.22094 0.321485 3
ACh/ AITC/ Hist/ Ment 0 0
ACh/ AITC/ Hist/ Cap 0 0.504697 0 1
ACh/ AITC/ Ment/ Cap 0 0
ACh/ ATP/ Hist/ Ment 0 0
ACh/ ATP/ Hist/ Cap 0.903462 0.167517 3 0.692863 0.257398 2 ACh/ ATP/ Ment/ Cap 1.148747 0 1 0 ACh/ Hist/ Ment/ Cap 0 0
AITC/ ATP/ Hist/ Ment 0 0
AITC/ ATP/ Hist/ Cap 1.459098 0 1 0 AITC/ ATP/ Ment/ Cap 0 0
AITC/ Hist/ Ment/ Cap 0 0
ATP/ Hist/ Ment/ Cap 0 1.894302 0 1
ACh/ AITC/ ATP/ Hist/ Ment 0 0
ACh/ AITC/ ATP/ Hist/ Cap 1.021866 0 1 1.501587 0 1 ACh/ AITC/ ATP/ Ment/ 0 0 Cap ACh/ AITC/ Hist/ Ment/ Cap 0 0
ACh/ ATP/ Hist/ Ment/ Cap 0 0
AITC/ ATP/ Hist/ Ment/ Cap 1.456125 0 1 0 The numbers are from two separate experiments for each treatment condition. Abbreviations: ATP, adenosine triphosphate; AITC, Allyl Isothiocyanate (mustard oil); KCl, potassium chloride.
Percent of DRG neurons responding to constellation pharmacology triggers Percentage of Cells Number of cells
0.002% 10 µM R9 CBD3 Trigger(s) 0.002% DMSO 10 µM R9 CBD3 A6K DMSO A6K responder Ach 25.33784 24.98078401 300 325 responder AITC 1.773649 1.229823213 21 16 responder ATP 17.48311 14.60415065 207 190 responder Histamine 3.040541 2.690238278 36 35 responder Menthol 5.405405 3.689469639 64 48 responder Capsaicin 33.19257 33.20522675 393 432 responder kcl 100 100 1184 1301 KCL only 34.62838 40.66102998 410 529 all triggers 0 0
ACh only 18.07432 16.21829362 214 211 AITC only 0.337838 0.307455803 4 4 ATP only 7.010135 4.842428901 83 63 Histamine only 0.337838 0.384319754 4 5 Menthol only 3.209459 1.767870869 38 23 Capsaicin only 20.77703 19.60030746 246 255 ACh/ AITC 0.084459 0
ACh/ ATP 2.111486 1.460415065 25 19 ACh/ Hist 0.422297 0.691775557 5 9 ACh/ Ment 0.337838 0.076863951
ACh/ Cap 2.364865 3.920061491 28 51 AITC/ ATP 0 0.076863951 1
AITC/ Hist 0.168919 0 2
AITC/ Ment 0 0.076863951 1
AITC/ Cap 0 0.153727902 2
ATP/ Hist 0.084459 0.153727902
ATP/ Ment 0.422297 0.076863951
ATP/ Cap 5.405405 5.611068409 64 73 Hist/ Ment 0.337838 0.230591852 4
Hist/ Cap 0.591216 0.230591852 7
Ment/ Cap 0.422297 0.614911606 8
ACh/ AITC/ ATP 0.084459 0.076863951
ACh/ AITC/ Hist 0.084459 0
ACh/ AITC/ Ment 0 0
ACh/ AITC/ Cap 0.084459 0 1
ACh/ ATP/ Hist 0 0.076863951
ACh/ ATP/ Ment 0 0
ACh/ ATP/ Cap 0.675676 1.306687164 8 17 ACh/ Hist/ Ment 0 0.230591852 3
ACh/ Hist/ Cap 0.253378 0.076863951
ACh/ Ment/ Cap 0.168919 0.153727902
AITC/ ATP/ Hist 0 0
AITC/ ATP/ Ment 0 0
AITC/ ATP/ Cap 0.506757 0 6
AITC/ Hist/ Ment 0 0
AITC/ Hist/ Cap 0 0
AITC/ Ment/ Cap 0.084459 0
ATP/ Hist/ Ment 0.168919 0.076863951
ATP/ Hist/ Cap 0.084459 0 1
ATP/ Ment/ Cap 0.084459 0 1
Hist/ Ment/ Cap 0 0.076863951
ACh/ AITC/ ATP/ Hist 0 0.076863951
ACh/ AITC/ ATP/ Ment 0 0.076863951
ACh/ AITC/ ATP/ Cap 0 0.230591852 3
ACh/ AITC/ Hist/ Ment 0 0
ACh/ AITC/ Hist/ Cap 0 0.076863951
ACh/ AITC/ Ment/ Cap 0 0
ACh/ ATP/ Hist/ Ment 0 0
ACh/ ATP/ Hist/ Cap 0.253378 0.153727902 3 2 ACh/ ATP/ Ment/ Cap 0.084459 0
ACh/ Hist/ Ment/ Cap 0 0
AITC/ ATP/ Hist/ Ment 0 0
AITC/ ATP/ Hist/ Cap 0.084459 0 1
AITC/ ATP/ Ment/ Cap 0 0
AITC/ Hist/ Ment/ Cap 0 0
ATP/ Hist/ Ment/ Cap 0 0.076863951
ACh/ AITC/ ATP/ Hist/ Ment 0 0
ACh/ AITC/ ATP/ Hist/ Cap 0.084459 0.076863951
ACh/ AITC/ ATP/ Ment/ Cap 0 0
ACh/ AITC/ Hist/ Ment/ Cap 0 0
ACh/ ATP/ Hist/ Ment/ Cap 0 0
AITC/ ATP/ Hist/ Ment/ Cap 0.084459 0 1
The numbers are from two separate experiments for each treatment condition. Abbreviations: ATP, adenosine triphosphate; AITC, Allyl Isothiocyanate (mustard oil); KCl, potassium chloride.
Frequency of DRG neurons per size responding to constellation pharmacology triggers 0.002% DMSO 10 µM R9 CBD3 A6K
499 599 649 699 849 1000 499 599 649 699 849 1000 399 399 - - 0 400 - 500 - 600 - 650 - 700 - 850 - >1000 0 400 - 500 - 600 - 650 - 700 - Trigger(s) 850 - >1000 responder Ach 71 49 52 18 15 45 21 29 59 69 55 37 24 33 25 23 responder AITC 9 6 1 1 0 1 1 2 3 3 3 1 2 2 1 1 responder ATP 73 42 36 13 10 17 7 9 61 40 34 13 11 14 7 10 responder 16 7 4 3 2 2 1 1 11 4 8 4 1 5 1 1 Histamine responder Menthol 28 8 8 2 3 7 5 3 12 7 12 8 0 6 2 1 responder 94 88 84 32 24 42 17 12 123 95 74 47 21 42 16 14 Capsaicin responder kcl 286 211 227 84 54 124 73 125 302 274 209 102 64 131 72 147 KCL only 78 63 75 30 14 35 34 81 107 108 75 22 19 57 36 105 all triggers 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ACh 52 36 38 14 9 31 14 20 37 48 33 22 17 22 13 19 AITC 2 2 0 0 0 0 0 0 0 0 1 0 1 1 0 1 ATP 27 18 17 5 4 7 1 4 24 14 8 4 4 2 1 6 Hist 1 2 1 0 0 0 0 0 0 0 3 0 1 1 0 0 Ment 14 4 6 1 3 5 3 2 4 6 7 2 0 2 2 0 Cap 49 52 63 23 14 26 11 8 77 57 37 30 12 25 8 9 ACh/ AITC 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 ACh/ ATP 4 2 3 1 1 5 5 4 0 5 4 1 1 3 4 1 ACh/ Hist 4 0 1 0 0 0 0 0 6 0 3 0 0 0 0 0 ACh/ Ment 1 2 0 0 0 0 0 1 0 0 0 1 0 0 0 0 ACh/ Cap 5 3 5 1 4 6 2 2 6 11 10 7 3 6 5 3 AITC/ ATP 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 AITC/ Hist 1 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 AITC/ Ment 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 AITC/ Cap 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 ATP/ Hist 1 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 ATP/ Ment 5 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 ATP/ Cap 26 15 11 5 2 4 0 1 26 16 18 3 3 5 0 2 Hist/ Ment 1 1 1 0 0 0 1 0 0 1 1 1 0 0 0 0 Hist/ Cap 2 1 1 0 1 1 0 1 1 1 1 0 0 0 0 0 Ment/ Cap 1 1 0 1 0 1 1 0 3 0 2 1 0 2 0 0 ACh/ AITC/ ATP 0 0 1 0 0 0 0 0 0 0 0 0 1 0 0 0 ACh/ AITC/ Hist 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ACh/ AITC/ Ment 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ACh/ AITC/ Cap 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ACh/ ATP/ Hist 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 ACh/ ATP/ Ment 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ACh/ ATP/ Cap 0 3 3 0 1 1 0 0 4 3 3 4 2 0 1 0 ACh/ Hist/ Ment 0 0 0 0 0 0 0 0 2 0 0 1 0 0 0 0 ACh/ Hist/ Cap 0 1 0 1 0 1 0 0 0 0 0 0 0 0 1 0 ACh/ Ment/ Cap 1 0 1 0 0 0 0 0 0 0 1 0 0 1 0 0 AITC/ ATP/ Hist 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 AITC/ ATP/ Ment 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 AITC/ ATP/ Cap 3 2 0 0 0 0 1 0 0 0 0 0 0 0 0 0 AITC/ Hist/ Ment 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 AITC/ Hist/ Cap 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 AITC/ Ment/ Cap 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 ATP/ Hist/ Ment 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 ATP/ Hist/ Cap 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 ATP/ Ment/ Cap 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Hist/ Ment/ Cap 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 ACh/ AITC/ ATP/ 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 Hist ACh/ AITC/ ATP/ 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 Ment ACh/ AITC/ ATP/ 0 0 0 0 0 0 0 0 1 0 1 0 0 0 1 0 Cap ACh/ AITC/ Hist/ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ment ACh/ AITC/ Hist/ 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 Cap ACh/ AITC/ Ment/ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Cap ACh/ ATP/ Hist/ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ment ACh/ ATP/ Hist/ 1 1 0 1 0 0 0 0 2 0 0 0 0 0 0 0 Cap ACh/ ATP/ Ment/ 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Cap ACh/ Hist/ Ment/ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Cap AITC/ ATP/ Hist/ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ment AITC/ ATP/ Hist/ 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Cap AITC/ ATP/ Ment/ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Cap AITC/ Hist/ Ment/ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Cap ATP/ Hist/ Ment/ 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 Cap ACh/ AITC/ ATP/ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Hist/ Ment ACh/ AITC/ ATP/ 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 Hist/ Cap ACh/ AITC/ ATP/ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ment/ Cap ACh/ AITC/ Hist/ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ment/ Cap ACh/ ATP/ Hist/ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ment/ Cap AITC/ ATP/ Hist/ 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ment/ Cap The numbers are from two separate experiments for each treatment condition. Abbreviations: ATP, adenosine triphosphate; AITC, Allyl Isothiocyanate (mustard oil); KCl, potassium chloride. The size bins (vertical text) are in µm2.
Supplementary Table 2
Mean % inhibition of control Receptor/Ion Channels Binding Assays ≥–50% ≤25% 25–50% ≥50% Receptors A1 adenosine receptor A2A adenosine receptor A3 adenosine receptor alpha 1 adrenergic receptor alpha 2 adrenergic receptor beta 1 adrenergic receptor beta 2 adrenergic receptor Angiotensin receptor 1 (AT1) Benzodiazepine receptor (central) Bradykinin receptor B2 Cannabinoid receptor 1 (CB1) Cholecystokinin receptor 1 (CCK1 or CCKA) Dopamine D1-like receptor Dopamine D2S receptor Endothelin receptor type A (ETA) Gamma aminobutyric acid receptor (GABAR) Galanin receptor 2 (GAL2) C-X-C chemokine receptor type 2 (CXCR2) Inflammatory chemokine receptor 1 (CCR1) Histamine receptor 1 (H1) Histamine receptor 2 (H2) Melanocortin Receptor 4 (MC4) Melatonin receptor 1 (MT1) Muscarinic receptor 1 (M1) Muscarinic receptor 2 (M2) Muscarinic receptor 3 (M3) Neurokinin receptor 2 (NK2) Neurokinin receptor 3 Neuropeptide Y receptor 1 (Y1) Neuropeptide Y Receptor (Y2) Neurotensin 1 receptor (NT1) delta 2 opioid receptor (δOR) kappa opioid receptor (κOP) Mu opioid receptor (µOP) Nociceptin receptor (NOP) aka opioid receptor-like (ORL1) Prostaglandin E receptor (EP4) Serotonin or 5-hydroxytryptamine (5-HT) receptor 1A (5- HT1A) 5-HT1B 5-HT2A 5-HT2B 5-HT5A 5-HT6 5-HT7 Somatostatin receptor (sst) vasoactive intestinal peptide (VIP) type 1 receptor (VPAC1) Vasopressin receptor 1A (V1a) Ion channels Benzodiazepine receptor Serotonin or 5-hydroxytryptamine (5-HT) receptor 3 (5-HT3) Ca2+ channel (L, verapamil site) Kv channel Small conductance Ca2+-activated K+ (SKCa) channel Na+ channel (site 2) Cl- channel (GABA-gated) Transporters norepinephrine transporter (h) dopamine transporter (h) Serotonin or 5-hydroxytryptamine (5-HT) transporter