Supplementary Table 1. Artery Nomenclature for Figure 4

Home , Cerebellar artery

(1) Superficial temporal (16) Profunda brachii Arteries in Inset (2) Facial (17) Ulnar collateral (1) Vertebral (3) External carotid (18) Radial branches (2) Basilar (4) Internal carotid (19) Coronary (3) Superior cerebellar (5) Common carotid (20) Intercostal (4) Hypophyseal portal (6) Vertebral (21) Common iliac (5) Anterior cerebral (7) Aortic arch (22) Femoral (6) Anterior communicating (8) Subclavian (23) Saphenous (7) Middle cerebral (9) Axillary (24) Iliaco-femoral (8) Internal carotid (10) Brachial (25) Superficial caudal epigastric (9) Posterior cerebral (11) Medial (26) Medial proximal genicular (10) Pontine (12) Radial (27) Popliteal (11) Anterior inferior cerebellar (13) Internal mammary (28) Proximal caudal femoral (12) Anterior spinal (14) Lateral thoracic (29) Gracilis (15) Subscapular (30) Median coccygeal

Supplementary Table 1. Artery nomenclature for Figure 4.

A B C D

3 mm 2 mm 3 mm 500 µm

E F G H

ICA ECA

500 µm 250 µm 250 µm 250 µm

I j J K L l

Brachial

500 µm 200 µm 200 µm 200 µm

M N O P

100 µm o 60 µm 60 µm 250 µm

Q R S T t r

300 µm 40 µm 300 µm 40 µm

Figure S1. Restricted TRPV1 expression in isolated aorta, skeletal muscle and mesenteric arteries. (A), nLacZ staining in the aortic arch and (B), descending aorta (showing TRPV1 expression is restricted to small feeding arteries “vasa vasorum”, see inset) and (C), abdominal aorta and (D), common carotid and (E), external (ECA) and internal carotid (ICA) arteries and (F), facial artery and (G), maxillary artery, and (H), superficial temporal artery and (I, J), axillary artery and ulnar collateral artery and (K, L),subscapular artery and branches and (M-O), muscle branches of brachial artery and (P), medial artery and branches and (Q-T), mesenteric arteries.

Radial branch A Axillary Brachial Medial Subscapular of brachial (#9) (#10) (#11) (#15) (#18)

100 µm

B Muscle artery of Proximal caudal femoral artery Radial branch of brachial Latisismus Dorsi (PCFA) and Gracilis artery (GA)

PCFA

40 µm

PCFA

40 µm 1 mm

Figure S2. TRPV1 expression in arteries of forelimb and skeletal muscle. (A) nLacZ staining (TRPV1PLAP-nlacZ mouse) and tdTomato fluorescence (TRPV1-Cre:tdTomato mouse) in forelimb trunk arteries, and (B) skeletal muscle arteries. Note that TRPV1 expression increases inversely proportional to arterial diameter (A) and with branching into tissue (B) and the correspondence of the nLacz staining and tdTomato signals.

PLAP staining nLacZ staining

A B C

Hypophyseal arteries

40 µm 120 µm 40 µm

D E F

E 100 µm 300 µm 100 µm Anterior interior cereballar artery G H I

H 100 µm 100 µm 100 µm Posterior interior cereballar artery

Figure. S3. TRPV1 expression in cerebral arteries. PLAP and nLacZ staining in (A to C), hypophyseal portal arteries, (D to F), anterior inferior cerebellar artery, and (G to I ), posterior inferior cerebellar artery.

A Control Cap (1 µM) HiK+ Cap HiK+

Wild-type B TRPV1-ChR2 60 µm TRPV1-Cre:tdTomato TRPV1-null WT Diameter (µm)

+ Cap HiK TRPV1-null Diameter (µm) ***

Diameter (%) *** Cap HiK+ *** ∆ TRPV1- Diameter (µm) Cre:tdTomato

Blue light Blue light Recovery Diameter (µm) TRPV1-ChR2 Time (s) C D E

Gracilis Diameter (µm) Diameter (µm)

Proximal caudal femoral 10-10 10-9 10-8 10-7 10-6 10-5 (M) 10-9 10-8 10-7 10-6 10-5 (M) Cap F Control Cap (1 µM) HiK+ G Cap HiK+ ) µm ( TRPV1 null Diameter 200 µm 200 µm Time (s)

Figure S4. Arterial contractility in response to TRPV1 agonist and optogenetic stimulation. (A, B), Responses of pressurized cerebellar branch arteries from WT, TRPV1-null, TRPV1-Cre:tdTomato and TRPV1-Cre:ChR2 to capsaicin and blue light (n = 4 - 7 arteries from 3 - 5 mice per group, ***P < 0.001). (C), nLacZ staining in gracilis artery of a TRPV1PLAP-nlacZ mouse. (D), Capsaicin-evoked constriction in isolated gracilis arteries with or without endothelium. (E), Acetylcholine (ACh) but not norepinephrine (NE) sensitivity is lost in denuded arteries. (F, H), Intravital imaging of a radial branch artery in a TRPV1-null mice in response to local application of capsaicin and KCl.

A B C D

*** ***

*** *** *** *** HR (bpm) SBP (mmHg) SBP DBP (mmHg) DBP MAP (mmHg) MAP

WT KO WT KO WT KO WT KO WT KO WT KO WT KO WT KO Cap LPA Cap LPA Cap LPA Cap LPA (32 µg/kg) (60 µg/kg) (32 µg/kg) (60 µg/kg) (32 µg/kg) (60 µg/kg) (32 µg/kg) (60 µg/kg)

E DRG F G H I MOUSE *** *** *** *** *** ***

200 µm HR (bpm) BP (mmHg) BP HR (bpm) BP (mmHg) BP Wild-type

DIA SYS MAP DIA SYS MAP Cap (32 µg/kg) Cap (32 µg/kg) Cap (32 µg/kg) Cap (32 µg/kg) 200 µm Anesthetized mice (nerve ablated) Conscious mice (nerve ablated)

K L M + atropine + atropine + atropine ** *** *** *** *** *** ** ** *** *** * *** *** ** * *** *** ** *** *** SBP (mmHg) SBP DBP (mmHg) DBP MAP (mmHg) MAP

2 4 8 16 32 2 4 8 16 32 2 4 8 16 32 2 4 8 16 32 2 4 8 16 32 2 4 8 16 32

RAT Cap (µg/kg) Cap (µg/kg) Cap (µg/kg) N O P Nerve ablated Nerve ablated + atropine * HR (bpm) HR (bpm) MAP (mmHg) MAP

2 4 8 16 32 2 4 8 16 32 2 4 8 16 32 2 4 8 16 32

Cap (µg/kg) Cap (µg/kg) Cap (µg/kg)

Figure S5. Effects of TRPV1 agonists and antagonists on systemic blood pressure. (A-D), Systolic BP, diastolic BP, mean arterial pressure and heart rate (HR) values in mice before, and the peak pressor response after, capsaicin administration. (E), X-gal staining in control DRG ganglia and slice. (F-I), BP and heart rate in anesthetized and conscious mice treated with RTX as neonates. (K-N) BP and heart rate responses (during peak pressor phase) to capsaicin in rats, [Note: capsaicin evokes similar responses with or without atropine pretreatment] (n = 6, *P < 0.05, ** P < 0.01, ***P < 0.001). (O, P), MAP and HR responses to capsaicin in sensory nerve-ablated rats (neonatal capsaicin treatment, n=5).

A B C Nerve ablated Nerve ablated Nerve ablated MAP (mmHg) MAP DBP (mmHg) DBP SBP (mmHg) SBP

1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 1 2 3 4 5 Application Cap (32 µg/kg) Application Cap (32 µg/kg) Application Cap (32 µg/kg) D Cap (32 µg/kg/min) TRPV1-null BP (mmHg) BP 5 min

Figure S6. BP responses to repeated capsaicin administration. (A-C), Diastolic (DBP), systolic (SBP) and mean arterial pressure (MAP) in WT and sensory nerve ablated mice during repeated administration of capsaicin (n=3). (D), Capsaicin infusion in TRPV1-null mouse does not alter BP.