FIGURE 10. mWake+ cells and scRNA-Seq populations in the VMH.
Representative images of mWake expression in the VMH are shown. (a) anti-V5 IF staining (green) in a mWake(V5/+) mouse. (b) Anti-V5 IF staining (green) with DAPI in a mWake(V5/+) mouse. (c) Chromogenic RNAscope ISH for mWake mRNA (red) with hematoxylin counterstain (purple) in wild-type (WT) mice. (d) Endogenous tdTomato fluorescence (red) in mWake(Cre/+) mice with DAPI. In (b-d), the dashed line denotes the boundary of the VMH. In (a), the scale bar denotes 200 μm. In (b), the scale bar is equivalent to 500 μm, and the arrows indicate the dorsal (d) and medial (m) directions, and both also apply to (c) and (d). (e) UMAP plot of all cells from Kim et. al. (2019) re-clustered. Clusters where putative mWake+ cells constitute >70% of the population are colored and represent 7 out of 46 identified transcriptional profiles: cluster 31 (Nup62cl, yellow), cluster 34 (Dlk1, Il1rapl2, steel blue), cluster 40 (Scgn, teal blue), cluster 42 (Nr5a1, Glipr1, dark purple), cluster 43 (Nr5a1, Tcf7l2, green), cluster 44 (Nr5a1, Ctxn3, light purple), cluster 45 (Nr5a1, Car8, deep purple). Abbreviations: 3v, third ventricle; VMH, ventromedial hypothalamus; and ZI, zona incerta.
