Figure 1
Targeting and RNA enrichment from LepRLH cells (A) LepR-Cre mice and wild type mice were injected in the LH with an AAV carrying Flex-L10:GFP (graphic of coronal section does not represent exact coordinates). In wild-type littermates (wt/wt) the GFP expression was very low (right image) compared to LepR-cre mice. In homozygous Cre (cre/cre) mice, 83.5% of the cells with immunohistochemically-detected GFP were in the LH (mean, n = 12 hypothalami). (B) Scatterplot of RNAseq results showing normalized counts from LepRLH fraction (IP) in x axis plotted against normalized counts from Hypothalamus fraction (IN) in y axis. Violet-colored dots represent genes that are enriched in IP (with log2FoldChange > 1.5 and adjusted p < 0.01) and dark grey-colored dots represent genes that are enriched in IN (with log2FoldChange < − 1.5 and adjusted p < 0.01). GFP, LepR and known marker genes of LepRLH cells are represented as dots colored in dark violet. (C) Scatterplot of RNAseq results showing genes de-enriched in IP fraction (enriched in IN fraction) as log2 of normalized counts from IN fraction (x-axis) plotted against Fold-enrichment (log2 of IP/IN normalized counts) (y-axis). Markers known to not be present in LepRLHcells (Agrp, Npy) and non-neuronal markers are represented as dark violet dots. (D) Barplot showing Fold-enrichment (log2 of IP/IN normalized counts) of GABAergic markers Slc32a1, Gad1 and Gad2 and glutamatergic marker Slc17a6 (mean ± SEM). IP: immunoprecipitated samples, IN: input samples.
