Fig. 4. Identification of LPS-activated cell types in the NTS–AP.
a, A uniform manifold approximation and projection (UMAP) plot of 7,824 and 4,443 neuronal nuclei from the NTS and the AP, respectively, from 20 mice. b, Expression of the indicated marker genes across identified clusters, with the colours of the clusters the same as in the UMAP in a. The size of the circle represents the fraction of cells in each cluster expressing the marker gene; the colour of the circle represents the minimum to maximum level of gene expression for each gene. The boxes on the left indicate the presence of the cluster in the AP or NTS. c, Method for TRAP2 labelling with LPS (0.5 mg kg−1, i.p.) in INTACT mice for subsequent processing for snRNA-seq. d, UMAP of TRAPed nuclei of excitatory (TRAP2VGLUT2, orange) or inhibitory (TRAP2VGAT, green) neurons superimposed on a UMAP of all neuronal nuclei from NTS–AP (grey). e,f, Percentage of TRAP2 neurons within each indicated cluster (excitatory, green; inhibitory, orange) from NTS (e) and AP (f). The heatmaps below the plots indicate log2[fold change] TRAPed nuclei for each cluster relative to the baseline sequencing in a. g–l, Activation of VGLUT2 neurons in the NTS–AP using activating (hM3Dq) DREADDs in VGLUT2–Cre mice. g, Cumulative food intake for 24 h post activation (saline, grey; CNO, cyan). h–j, Food intake (h, paired t-test P = 8.3 × 10−7), water intake (I, paired t-test P < 1 × 10−7) and movement (j, paired t-test P = 4.1 × 10−5) at 12 h post activation of saline-treated (light cyan) or CNO-treated (dark cyan) animals. k,l, Change in core temperature (paired t-test P = 8.6 × 10−4) 4 h after (k) and percentage body weight (paired t-test P = 8.0 × 10−5) 24 h after (l) activation of saline-treated (light cyan) or CNO-treated (dark cyan) animals. g–l, Saline-treated animals, N = 7; CNO-treated animals, n = 7. ***P < 0.001, ****P < 0.0001. All error bars represent the s.e.m. The mouse illustration in c was created using BioRender.com.