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. 2022 Aug 31;609(7927):569–574. doi: 10.1038/s41586-022-05137-7

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a,b, Comparison of ROOT and AAV9 for retrograde labelling in the iWAT. a, Representative whole-mount images of DRGs (T13) and SChGs (T12) labelled by ROOT-mScarlet or AAV9-mScarlet and a sequential CTB-647 injection from iWAT. Some of the AAV and CTB double-positive cells are highlighted by white arrows. CTB labels 26.42 ± 6.49% of ROOT-labelled neurons, and 9.95 ± 0.92% of AAV9-labelled neurons in T13/L1 DRGs. Data are mean ± s.e.m. n = 4 mice per group. b, Representative images of livers from animals with ROOT-mScarlet or AAV9-mScarlet injected into the iWAT. c–f, Combinatorial viral strategy for Cre-dependent ablation of iWAT DRGs. c, Schematic of the combinatorial viral strategy for unilateral sensory ablation. Each mouse has AAV-mCherry-flex-DTA injected into the bilateral T13/L1 DRGs, and ROOT-YFP or ROOT-Cre injected into the iWAT unilaterally. Three weeks after surgery, CTB-647 was injected into the iWAT bilaterally. d, Representative whole-mount images of contralateral (Cre⁻) and ipsilateral (Cre⁺) DRGs (T13) and SChGs (T12). e,f, Quantification of CTB⁺ cell numbers (e) and normalized cell numbers (f) in T13 and L1 DRGs labelled from iWAT. n = 10 mice. Statistical analysis was performed using two-tailed paired t-tests. P values are shown at the top. For a,b and d, scale bars, 200 μm.

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