Interleukin-17 Regulates Neuron-Glial …...Figure S6 A schematic showing how IL-17 modulates excitatory synaptic transmission (a) and inhibitory synaptic transmission (b) in spinal

Cell Reports, Volume 29

Supplemental Information

Interleukin-17 Regulates Neuron-Glial

Communications, Synaptic Transmission,

and Neuropathic Pain after Chemotherapy

Hao Luo, Hui-Zhu Liu, Wen-Wen Zhang, Megumi Matsuda, Ning Lv, Gang Chen, Zhen-Zhong Xu, and Yu-Qiu Zhang

Supplemental Figures

Figure S1 Co-localization of IL-17R and SOM+ neurons in lamina II superficial spinal cord dorsal horn. Related to Figure

2 and 3. (A) RNAsope ISH showing co-localization of IL-17R and SOM/SST mRNAs in lamina II superficial spinal cord dorsal

horn neurons. Yellow arrows should double-labeled neurons. After ISH, the spinal sections were co-labeled with anti-GFAP

antibody or DAPI. Note IL-17R mRNA expression is low but highly specific. There are no positive white puncta outside the

spinal cord tissue shown by star. Scale, 20 mm. (B) Immunofluorescence staining of IL-17R-IR (blue) with SOM+ neurons

(red) expressing IL-17R-shRNA-EGFP (green) in spinal dorsal horn. The specificity of IL-17R knockdown was verified by

overlap of IL-17R-shRNA-EGFP (green), IL-17R-IR (blue) and SOM-tdTomato (red) signals. No IL-17R-IR (arrow) was detected

in IL-17-shRNA-EGFP expressing SOM+ neurons (arrow head). Scale bar: 50 μm.

Figure S2 Colocalization of IL-17 (green) with tdTomato (red) in spinal cord of IL-17-tdTomato mouse. Related to Figure

4. The specificity of IL-17 antibody was verified by extended overlap of IHC and IL-17-tdTomato signals (arrow heads) The box

in the up panel was enlarged in the bottom panel. Scale bar: 50 μm (top) and 20 μm (bottom).

Figure S3 IL-17 expression in spinal cord dorsal horn. Related to Figure 4. Double staining of IL-17 with IB4 (left) and

CGRP (right) in the spinal cord dorsal horn. The boxes in the up panels were enlarged in the bottom panels. Scale bar: 50 μm (top)

and 20μm (bottom).

Figure S4 IL-17 expression in DRG. Related to Figure 5. (A) Colocalization of IL-17 (red) with satellite glial marker glutamine

synthetase (GS, green). Scale bar: 50 μm. The inserts are enlarged images. Scale bar: 10 μm. (B) Colocalization of IL-17-GFP

(red) with satellite glial marker glutamine synthetase (GS, green) in IL-17-GFP mice.

Figure S5 Double staining of IL-17 with the T cell marker CD3 in spinal dorsal horn. Related to Figure 7. Note there is no

colocalization between IL-17 and CD3. Scale bar: 50 μm (top) and 20 μm (bottom).

Figure S6 A schematic showing how IL-17 modulates excitatory synaptic transmission (a) and inhibitory synaptic

transmission (b) in spinal SOM+ neurons to induce central sensitization and pain hypersensitivity. Related to Figure 1,

Figure 2 and discussion. (a) IL-17 is released from activated astrocytes and acts on IL-17R on presynaptic terminals and

postsynaptic SOM+. Activation of presynaptic IL-17R results in increased glutamate release, whereas activation of postsynaptic

IL-17R also enhances NMDAR activity. (b) IL-17 is also expressed by inhibitory neurons and facilitates GABA release to inhibit

an inhibitory neuron, leading to a reduction in presynaptic inhibitory control of spinal SOM+ neurons. This disinhibition will also

cause hyperactivity in SOM+ neurons.

Figure S7 A schematic of IL-17-induced neuron-glial interaction in DRG. Related to Figure 5, Figure 6 and discussion. IL-

17 is expressed by satellite glial cells. Following chemotherapy, IL-17 released from satellite glia acts on IL-17R on nociceptive

neurons to increase neuronal excitability and induce peripheral sensitization.