[https://doi.org/10.5061/dryad.9s4mw6msg](https://doi.org/10.5061/dryad.9s4mw6msg)

## Description of the data and file structure

We provide raw data of main figures 1 to 4 organized by figure number.

Figure 1

Fig 1C: Number of tissue Tregs across organs, in both sexes combined. Treg numbers were normalized to organ weight (g).

Fig 1D: Number of tissue Tregs from male and female mice across organs. Treg numbers were normalized to female Treg numbers.

Fig 1F: Number of tissue Tregs 2 days after a single IT pegDT injection across organs in female mice. Treg numbers were normalized to vehicle-injected Treg numbers.

15 represents mean number of tissue Tregs in IT vehicle-injected mice per organ.

Fig 1G: Nociceptive thresholds following mTreg depletion in female and male mice.

Fig 1H: Behavioral differences between control and mTreg depleted mice in female and male mice.

Figure 2

Fig 2C: Percent response to 0.008 g von Frey filament in mTreg depleted female mice with SNI.

Fig 2D: Percent response to 0.008 g von Frey filament in mTreg depleted male mice with SNI.

Fig 2F: Number of tissue Tregs in the spinal cord meninges following mTreg expansion using IL-2, in both sexes combined. Treg numbers were normalized to organ weight (g).

Fig 2G: Nociceptive thresholds following mTreg expansion in female mice with SNI.

Fig 2E: Nociceptive thresholds following mTreg expansion in male mice with SNI.

Fig 2J: Nociceptive thresholds following mTreg depletion in uninjured Four Core Genotype female and male mice.

Fig 2K: Percent baseline nociceptive thresholds of the data in Fig. 2J.

Fig 2L: Nociceptive thresholds following mTreg expansion in Four Core Genotype female mice with SNI.

Fig 2M: Nociceptive thresholds following mTreg expansion in Four Core Genotype male mice with SNI.

Fig 2N: Anti-nociceptive efficacy of the data in Fig. 2L and M.

Fig 2P: Nociceptive thresholds following mTreg expansion in ovariectomized female mice with SNI injected with estrogen or progesterone or estrogen antagonist.

Fig 2Q: Anti-nociceptive efficacy of the data in Fig. 2P.

Figure 3

Fig 3F: Number of tissue Tregs across organs in the spinal cord meninges in female and male mice following mTreg expansion using IL-2. Treg numbers were normalized to organ weight (g).

Fig 3G: Enkephalin levels in mouse CSF following mTreg depletion or exapansion.Enkephalin levels were normalized to CSF volume collected (ml).

Fig 3H: Nociceptive thresholds following mTreg depletion in uninjured female PenkCreRosa26DTR bone marrow chimera mice.

Fig 3I: Nociceptive thresholds following mTreg depletion in uninjured male PenkCreRosa26DTR bone marrow chimera mice.

Fig 3J: Nociceptive thresholds following mTreg depletion in nerve injured female PenkCreRosa26DTR bone marrow chimera mice.

Fig 3K: Nociceptive thresholds following mTreg depletion in nerve injured male PenkCreRosa26DTR bone marrow chimera mice.

Fig 3L: Nociceptive thresholds following Treg specific enkephalin depletion in uninjured female Foxp3CreERT2;Penkfl mice.

Fig 3M: Nociceptive thresholds following Treg specific enkephalin depletion in nerve injured female Foxp3CreERT2;Penkfl mice.

Fig 3N: Nociceptive thresholds following mTreg specific enkephalin depletion in uninjured female Foxp3-DTR:PenkCreRosa26DTR mixed marrow chimera mice.

Fig 3O: Nociceptive thresholds following mTreg specific enkephalin depletion in nerve injured female Foxp3-DTR:PenkCreRosa26DTR mixed marrow chimera mice.

Figure 4

Fig 4b: Anti-nociceptive efficacy of mTreg expansion following naltrindole IT injection in nerve injured female mice.

Fig 4C: Nociceptive thresholds and anti-nociceptive efficacy following mTreg expansion in Oprd1 flox female mice with SNI injected with AAV-Cre targeting the PNS.

Fig 4D: Nociceptive thresholds following mTreg expansion in Oprd1 flox female mice with SNI injected with AAV-Cre targeting the CNS.

Fig 4E: Expression from DRG sensory neurons clusters from combined GSE139088 and GSE201653. Expression was normalized by row.

Fig 4K: Area under the curve of DRG neuron calcium imaging following von Frey stimulation following mTreg expansion in nerve injured MrgprdDCre-ERT2;GCaMP6f female mice.

Fig 4L: Percent response to 0.008 g von Frey fiber stimulation after SNI in MrgprD-CreERT2;Oprd1fl female mice.

Fig 4M: Nocifensive behaviors following 0.008 g von Frey fiber stimulation after SNI in MrgprD-CreERT2;Oprd1fl female mice.

Fig 4N: Anti-nociceptive efficacy of Deltorphin IT injection in nerve injured MrgprD-CreERT2;Oprd1fl female mice. LMC: Littermate controls

Fig 4O: Nociceptive thresholds following mTreg expansion in nerve injured MrgprD-CreERT2;Oprd1fl female mice.

Fig 4P: Anti-nociceptive efficacy of mTreg expansion in MrgprD-CreERT2;Oprd1fl female mice.LMC: Littermate controls

Experimental data files of main figures. Each .xls contains main figure files delineated by individual sheets corresponding to a subpanel. 

Analyzed data file

DRG data.xls includes analyzed data set showing the number of neurons with* Oprd1* expression of >1 from Figure 4E-G. 

There is an analyzed normalization of RNA expression amongst activated, resting Tconv and Treg from Figure 3A-B. 

There are 4 ATAC-seq bedwig files of concocted samples of activated, resting Tconv and Treg from Figure 3C. 

## Acknowledgements

We would like to acknowledge the following prior work and publicly available datasets from:

J. van der Veeken, A. Glasner, Y. Zhong, W. Hu, Z.-M. Wang, R. Bou-Puerto, L.-M. Charbonnier, T. A. Chatila, C. S. Leslie, A. Y. Rudensky, The transcription factor Foxp3 shapes regulatory T cell identity by tuning the activity of trans-acting intermediaries. Immunity 53, 971-984.e5 (2020).

A. Baysoy, K. Seddu, T. Salloum, C. A. Dawson, J. J. Lee, L. Yang, S. Gal-oz, H. Ner-Gaon, J. Tellier, A. Millan, A. Sasse, B. Brown, L. L. Lanier, T. Shay, S. Nutt, D. Dwyer, C. Benoist, The Immunological Genome Project Consortium, The interweaved signatures of common-gamma-chain cytokines across immunologic lineages. Journal of Experimental Medicine 220, e20222052 (2023). Specifically, https://www.immgen.org/

S. A. Bhuiyan, M. Xu, L. Yang, E. Semizoglou, P. Bhatia, K. I. Pantaleo, I. Tochitsky, A. Jain, B. Erdogan, S. Blair, V. Cat, J. M. Mwirigi, I. Sankaranarayanan, D. Tavares-Ferreira, U. Green, L. A. McIlvried, B. A. Copits, Z. Bertels, J. S. Del Rosario, A. J. Widman, R. A. Slivicki, J. Yi, R. Sharif-Naeini, C. J. Woolf, J. K. Lennerz, J. L. Whited, T. J. Price,  null Robert W Gereau Iv, W. Renthal, Harmonized cross-species cell atlases of trigeminal and dorsal root ganglia. Sci Adv 10, eadj9173 (2024). https://harmonized.painseq.com/