Figure 2.

PMv and Arc LepRb Cell-Specific Transcriptional Changes Induced by Short-Term Leptin Treatment

(A) LepRb eGFP-L10a mice were divided into three groups: diestrous females treated with i.p. saline (WT) and Lep^ob females treated with i.p. saline (Lep^ob) or i.p. leptin (Lep^ob + leptin). Mice were euthanized 1 h after the last saline or leptin injections. PMv (left and right sides) and Arc (median line) were collected by micro punches (1.25 mm diameter) of individual mouse brains and LepRb_TRAP genes were subjected to RNA-seq. GFP fluorescence (green) shows the dense distribution of LepRb neurons in the PMv and Arc.

(B) Enrichment fold of Lepr in individual samples was used as control.

(C) Heatmap showing DEGs across treatment groups.

(D, F, and G) Venn diagrams showing number of DEGs comparing treatment groups in LepRb PMv (D) and Arc (G) neurons, and the number of recovered DEGs (rDEGs) in both nuclei (F).

(E and H) Enriched rDEGs in PMv (E) and Arc (H).

(I) Single (a and b) and dual (c and d) label immunofluorescence showing VWF immunoreactivity in endothelia of blood vessels (bv) in the brain parenchyma (a–c) and in the base of the brain (d). Blood vessels were labeled with anti-laminin in red, and VWF-ir was labeled in green (in a, c, and d). In b, GFP autofluorescence was visualized in green and VWF-ir in red. Arrow indicates a LepRb eGFP-L10a cell (GFP autofluorescence).

(J) rDEGs in both nuclei.

Scale bars: 400 μm in (A); 200 μm in (Ia), and 50 μm in (Ib, Ic, and Id). See Figures S2–S4 and Tables S2 and S3.