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. 2019 Aug 30;16(7):1200–1220. doi: 10.1080/15548627.2019.1659616

Figure 7.

Figure 7.

Intracerebroventricular (ICV) injection of PK11195 or ablation of Tspo induces lipophagy in tanycytes. (A) Representative micrographs and quantification of colocalization for p-PRKAA (red) and VIM (green) in ventral tanycytes of PK11195- or vehicle-injected mice fed NCD (n = 18 sections/3 mice). (B) Immunoblotting analysis and quantification of p-PRKAA and autophagy markers in the hypothalamus after ICV injection of PK11195 (PK), compound C (CC), or vehicle into the 3V of mice fed NCD (n = 6). (C) Immunostaining for LAMP2 (red) and VIM (magenta) with BODIPY (green) and Hoechst (blue) staining in ventral tanycytes of mice fed NCD with or without injection of PK11195 or OL, or fed HFD with or without PK11195. Average number and size of LDs per cell and colocalization of BODIPY and LAMP2 (white) were quantified (n = 15 sections/3 mice). (D–F) Tspofl/fl mice were injected with AAV-Rax-Cre-GFP or AAV-Rax-GFP and fed HFD for 4 weeks. (D and E) Immunostaining for p-PRKAA (red; D) or MAP1LC3B (red; E), and VIM (magenta) in GFP+-tanycytes. Colocalization of p-PRKAA and VIM (white) was quantified (D; n = 9 sections/3 mice). Aaverage number of MAP1LC3B puncta in the cytosol (arrowhead) was also quantified (E; n = 58–72 cells/3 mice). (F) Immunostaining for TSPO (red) and VIM (magenta) with BODIPY (green) and Hoechst (blue) staining in ventral tanycytes. Average number and size of LDs per cell were quantified (n = 88 or 89 cells/3 mice). Data are mean ± SEM, *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 vs. PK11195, (NCD) vehicle, or AAV-Rax-GFP; ###P ≤ 0.001 vs. (NCD) OL; ††P ≤ 0.01, †††P ≤ 0.001 vs. (HFD) vehicle, as determined by Student’s t-test.