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. 2021 Jan 26;47:101172. doi: 10.1016/j.molmet.2021.101172

Figure 2.

Figure 2

Analysis of hedgehog signaling in Glast+astrocytes after Patched deletion. (A) mRNA levels of Gli1, Gli2, and Gli3 measured by qRT-PCR in the hypothalamus and cerebral cortex of the YFP-Ptc+/+ and YFP-Ptc−/− mice 4 weeks after tamoxifen (Tx). Actin served as a housekeeping gene for relative mRNA expression levels (n = 4 mice). (B) RNAscope for Gli1 and counterstaining for DAPI on coronal sections of the tuberal region of the hypothalamus highlighting the ventromedial hypothalamic nuclei and cerebral cortex of the YFP-Ptc+/+ and YFP-Ptc−/− mice 10 days after Tx. Insets highlight the difference in RNAscope signal intensity between the hypothalamus and cerebral cortex at the level of a single cell. (C) Quantitative analysis of RNAscope signals for Gli1 mRNA in Glast+ and S100β+ cells in hypothalamic nuclei and cerebral cortex from the YFP-Ptc+/+ and YFP-Ptc−/− mice. Bar graphs (A and C) represent mean ± SEM. n = 3–4 mice/group, ∗p < 0.05 by the Mann–Whitney test. Staining was replicated on three mice. Scale bars, 50 μm in (B). ARC, arcuate nucleus; VMH, ventromedial hypothalamic nucleus; DMH, dorsomedial hypothalamic nucleus.