Fig. 4

Pharmacological inhibition of calcineurin decreases GFAP fluorescence intensity in the ARC of mice acutely exposed to HFD. WT mice on chow (a, b) and HFD (c, d) were treated with 1 mg/kg/day of calcineurin inhibitor Fk506 for 1 week; HFD feeding was started in parallel. Fk506 treatment resulted in a significant reduction in body weight in mice on chow (a) and HFD (c), while no significant changes in body composition on chow (b) and HFD (d) were detected. GFAP (e, f) and IBA1 (g, h) staining in the ARC, VMH, and DMH of vehicle and Fk506-treated WT mice on HFD (scale 200 μm) revealed a decrease in GFAP fluorescence intensity in the ARC (e, f)

Real-time qPCRs of mice on chow and HFD (i–m) were performed for Gfap and Iba1 (i), inflammatory cytokines (j), Cd169 as marker for monocyte infiltration (k), calcineurin subunits (l), and calcineurin downstream targets (m). Means ± SEM; n = 6 biological replicates (a, b); n = 10 biological replicates, except for lean mass vehicle, where one value was identified as a significant outlier (c, d); n = 4 biological replicates (f, g); n = 6 biological replicates, except for Gfap chow vehicle, where one value was identified as a significant outlier (i–m)