Fig 2: Effect of ANA-12 on the L7Gn-induced upregulation of BDNF, p-TrkB, p-CREB, and p-ERK in the hippocampus of sleep-deprived mice. L7Gn was dissolved in 70% PEG-saline, and ANA-12 was dissolved in 1% DMSO. Mice were administered vehicle (70% PEG), L7Gn (3 mg/kg) or ANA-12 (0.5 mg/kg) once daily for five days. Western blotting and quantification of expression levels of (A) BDNF, (B) p-TrkB, (C) p-ERK, and (D) p-CREB. The expression level of BDNF was normalized against GAPDH and those of p-TrkB, p-ERK, and p-CREB were normalized against their unphosphorylated form. (A) BDNF: (one-way ANOVA: F (3, 12) = 31.64, p < 0.0001) (B) p-TrkB: (one-way ANOVA: F (3, 12) = 12.3, p = 0.0006) (C) p-ERK: (one-way ANOVA: F (3, 11) = 10.97, p = 0.0012) (D) p-CREB: (one-way ANOVA: F (3, 12) = 12.11, p = 0.0006). Data are presented as mean ± SEM (n = 3–4/group). * p < 0.05, significantly different from the control group. # p < 0.05, significantly different from the vehicle group. $ p < 0.05, significantly different from the L7Gn (3 mg/kg) group. CTL, control; Veh, vehicle.

Fig 3: Representative western blots of BDNF, oxytocin and vasopressin receptors.The figure shows representative western blots of TrkB (BDNF receptor), OT-R (oxytocin receptor), V1a, V3 (vasopressin receptors) and GAPDH (for normalization) with the corresponding molecular weight (expressed in kDa), in the prefrontal cortex, striatum, hypothalamus and hippocampus of mice belonging to emotional contagion resistant (ECR) and emotional contagion prone (ECP) subgroups.

Fig 4: Molecular targets of 7,8-DHF, quercetin, and apigenin in SH-SY5Y cells expressing ΔK280 TauRD-DsRed. Congo red (10 μM) was included as a negative control. Relative TRKB, p-TRKB (Y817), p-TRKB/TRKB, extracellular signal-regulated kinase (ERK), p-ERK (T202/Y204), cAMP-response element binding protein (CREB), p-CREB (S133), BCL2 apoptosis regulator (BCL2), and BCL2-associated X protein (BAX) protein levels were analyzed through immunoblotting using specific antibodies (n = 3). GAPDH was used as a loading control. Relative protein levels are shown on the right side of the representative western blot images. To normalize, the relative protein level in uninduced cells (Dox -) was set as 100%. p-values: comparisons between induced and uninduced cells (#p < 0.05) or between treated and untreated cells (*p < 0.05, **p < 0.01, ***p < 0.001) (one-way ANOVA with the post hoc Tukey’s test).

Fig 5: Pharmacological TrkB activation induced RBM3 without cooling and is neuroprotective in prion-diseased mice.(A) Schematic showing experimental design. Prion-diseased tg37 mice were treated with the TrkB agonist 7,8-DHF from 3 w.p.i. onwards. (B) Western blots showing sustained high levels of RBM3 and RTN3 after 7 wk of treatment at 10 w.p.i. Bar charts show mean ± SEM quantification. t test; **P < 0.01 RBM3, prion 10 w.p.i. + vehicle versus prion 10 w.p.i. + 7,8-DHF (P = 0.0075); RTN3, prion 10 w.p.i. + vehicle versus prion 10 w.p.i. + 7,8-DHF (P = 0.0089). n = 4 mice per condition. (C) Representative electron microscopy micrographs showing synapses in the CA1 region of the hippocampus of prion-diseased mice at 7 w.p.i. Presynaptic compartment pseudo-colored in yellow, post-synaptic compartment in green. 7,8-DHF rescued synapse loss in prion-diseased mice (compare panels b and c). Bar chart shows mean ± SEM quantification from n = 3 animals (93 images) per condition. Two-way ANOVA test; *P < 0.05, **P < 0.01 normal brain homogenate (NBH) control versus prion 7 w.p.i. + vehicle (P = 0.0134); prion 7 w.p.i. + vehicle versus prion 7 w.p.i. + 7,8-DHF (P = 0.008). Scale bar, 1 μm. (D) Novel object recognition deficits in prion-diseased mice are rescued by 7,8-DHF treatment at 9 and 10 w.p.i. Kruskal–Wallis test and Dunn’s multiple comparisons test, *P < 0.05, NBH control versus prion 9 w.p.i. (P = 0.0241); prion 9 w.p.i versus prion 9 w.p.i + 7,8-DHF (P = 0.0470); prion 9 w.p.i versus vehicle prion mice 10 w.p.i. (P = 0.0254). n = 8 control mice, 9 prion + vehicle mice, and 10 prion + 7,8-DHF treated mice. (E) Hematoxylin and eosin stained sections showing the CA1 region of the hippocampus (left). 7,8-DHF rescued pyramidal neuronal loss and reduced spongiosis in prion-diseased mice at terminal time point (12 w.p.i.) (compare panels b and c) and see bar chart (right) showing quantification of pyramidal neuronal counts (obtained by NeuN staining). One-way ANOVA test and Tukey’s multiple comparisons test, *P < 0.05 NBH control versus prion + vehicle (P = 0.0325; prion 9 w.p.i versus prion 9 w.p.i) + 7,8-DHF (P = 0.0119). Scale bar, 50 μm. n = 3–4 mice per condition.