Figure 5

Transcriptomic profiling of hypothalamus identifies genes differentially regulated by EPC in WT and KO mice. (A) Heat map of 101 differentially expressed transcripts identified by RNAseq. N = 3 mice per group. (B) Venn diagram showing the number of differentially expressed genes in KO and WT hypothalamus after EPC. Arrows indicate direction of change after EPC. (C) RT-qPCR confirmed altered expression of 5 genes of interest in the hypothalamus. (D) Representative immunoblots of hypothalamic protein extracts shows reduced expression of Ptgds and Gfap in WT mice after EPC (WT-H) compared to controls (WT-NH). The opposite change was observed in KO. Blots were quantified and normalized to actin, expressed as fold change relative to NH-WT. N = 4 mice per group. *p < 0.05 by unpaired t-test. (E) ELISA assay of PGD2 levels in hypothalamus. N = 4 mice per group. *p < 0.05 by unpaired t-test. Two-way ANOVA revealed a significant interaction between genotype and treatment on PGD2 levels (F = 32.3, df:1, p = 0.00046).