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. 2020 May 21;39:101022. doi: 10.1016/j.molmet.2020.101022

Figure 3.

Figure 3

Glial deletion of Nemo (NemogliaKO) interferes with the induction of NF-κB target genes by IL-1β. (A–I) Immunohistochemistry showed that IL-1β (20 μg/kg, i.v.) upregulated the NF-κB target gene VCAM1 in α-tanycytes of NemoFL but not of NemogliaKO mice. Representative immunostainings of VCAM1 in coronal mediobasal hypothalamus (MBH) sections of NemoFL (A–C) and NemogliaKO (D–F) mice 8 h after IL-1β treatment and PBS-treated NemoFL mice (G–I). (B), (C); (E), (F); and (H), (I) are higher magnifications of the boxed areas in (A), (D), and (G), respectively. Scale bar, 100 μm. (J) Quantification of VCAM1 staining in α-tanycytes projecting mainly into the VMH at various time points after IL-1β administration. Values are means ± SEM. Two-way ANOVA for genotype, F(1, 29) = 4.46, P = 0.044. ∗P < 0.05 (Bonferroni posttest, n = 7–8 animals per group). (K) Nissl-stained coronal section of the MBH showing microdissected areas of the tanycytic layer and arcuate nucleus (ARC). ME, median eminence; 3 V, 3rd ventricle; scale bar, 100 μm. (L) Representative agarose gels after RT-PCR for Nemo and β-actin in microdissected tanycytes from NemoFL and NemogliaKO mice. (M, N) Ptgs2 mRNA was reduced in tanycytes of NemogliaKO mice in comparison to NemoFL controls 4 h after IL-1β treatment (M). In contrast, there was no difference in the ARC (N). ∗P < 0.05 (Mann–Whitney test, n = 6–8 mice/group). (O) Pomc mRNA was reduced in tanycytes of NemogliaKO mice in comparison to NemoFL controls 4 h after IL-1β treatment (Mann–Whitney test, n = 6–8 mice/group). (P, Q) Plasma concentrations of IL-6 (P) and IL-1β (Q) did not differ between NemoFL and NemogliaKO animals after intravenous IL-1β injection. Values are means ± SEM (n = 6–8 mice/group).