Extended Data Fig. 2: Y1R co-expression with previously identified pain-related neuron populations in the PBN.
From: A parabrachial hub for need-state control of enduring pain
(a) UMAP embedding and representative spatial maps showing normalized expression levels of Pdyn, Tac1, and Oprm1. UMAP scale maximum: Pdyn, 2.7; Tac1, 2.55; Oprm1, 2.3. Spatial scale maximum: Pdyn, 1.9; Tac1, 1.9; Oprm1, 1.8. (b) UMAP embedding and representative spatial maps showing normalized expression levels of the IEGs Arc, Egr1, and Jun. UMAP scale maximum: Arc, 3; Egr1, 2.4; Jun, 2.75. Spatial scale maximum: Arc, 1.75; Egr1, 2; Jun, 2. (c) Diagram depicting 4 subregions of the lPBN. (d) Representative image showing RNAscope fluorescence in situ hybridization (FISH) used to validate CosMx results. Npy1r expression was observed throughout the PBN. Scale bar, 200 μm. (e) % of neurons expressing Npy1r in each subregions of the lPBN. Each dot represents one section (n = 3 mice). (f) Overlap between Calca and Npy1r expression from CosMx analysis. (g) Representative image showing Calca and Npy1r in the lPBN labeled using RNAscope FISH. Scale bar, 100 μm; inset, 50 μm. (h) % of Npy1r-expressing neurons co-expressing Calca and % of Calca-expressing neurons co-expressing Npy1r. Each dot represents one section (n = 3 mice). (i) Overlap between Pdyn and Npy1r expression from CosMx analysis. (j) Representative image showing Pdyn and Npy1r in the lPBN labeled using RNAscope FISH. Scale bar, 100 μm; inset, 50 μm. (k) % of Npy1r-expressing neurons co-expressing Pdyn and % of Pdyn-expressing neurons co-expressing Npy1r. Each dot represents one section (n = 3 mice). (l) Overlap between Penk and Npy1r expression from CosMx analysis. (m) Representative image showing Penk and Npy1r in the lPBN labeled using RNAscope FISH. Scale bar, 100 μm; inset, 50 μm. (n) % of Npy1r-expressing neurons co-expressing Penk and % of Penk-expressing neurons co-expressing Npy1r. Each dot represents one section (n = 4 mice). (o) Overlap between Tacr1 and Npy1r expression from CosMx analysis. (p) Representative image showing Npy1r and Tacr1 in the lPBN labeled using RNAscope FISH. Scale bar, 100 μm; inset, 50 μm. (q) % of Npy1r-expressing neurons co-expressing Tacr1 and % of Tacr1-expressing neurons co-expressing Npy1r. Each dot represents one section (n = 3 mice). Data are expressed as mean ± SEM.
