Fig. 5. Divalent cations downregulate sympathetic activity through hypothalamic CREB signaling.

Representative immunofluorescent images (a) and corresponding quantification (b) showing the phosphorylation of CREB in the ventromedial hypothalamus (VMH) 1 week postoperatively (n = 4). Lower images (scale bars = 20 µm) are high-resolution versions of the boxed regions in the upper images (scale bars = 200 µm). Representative Western blots (c) and corresponding quantification (d) showing the phosphorylation of CREB and the expression of 5-hydroxytryptamine receptor 2C (HTR2C) in the hypothalamus tissue of mice at week 1 postoperatively. e ELISA of serum and urine epinephrine levels of mice at week 1 postoperatively. f Representative immunofluorescent images showing the presence of tyrosine-hydroxylase⁺ (TH⁺) sympathetic fibers on cortical bone surface (n = 3, scale bars = 50 µm). g The daily distance (m), duration (s), and maximum speed of running-wheel activity of WT mice at weeks 1 and 4 postoperatively (n = 5). Representative immunofluorescent images showing the phosphorylation of CREB in the VMH of TrkA^wt or TrkA_Avil^−/− mice (h), as well as COX2^wt or COX2_LysM^−/− mice (j) at week 1 postoperatively (n = 3). Lower images (scale bars = 20 µm) are high-resolution versions of VMH regions in the upper images (scale bars = 200 µm). The daily distance (m) and duration (s) of running-wheel activity of TrkA^wt mice or TrkA_Avil^−/− (i), as well as COX2^wt or COX2_LysM^−/− (k) mice at week 4 postoperatively. Data are mean ± s.d. *P < 0.05, **P < 0.01 by Student’s two-sided T-test (d, e), two-way ANOVA with Tukey’s post hoc test (g), or 1-way ANOVA with Tukey’s post hoc test (b, i, k). Source data are provided as a source data file.