It is well established that leptin binding to ObRb activates JAK-STAT signalling so to confirm blockade of leptin binding to ObRb we monitored Stat-3 phosphorylation.
As an additional control, a Myc antibody (9E10) was able to reduce pathology when h1-120myc α-syn pffs were used, confirming blockade of exogenous pff uptake into neurons.
The specificity of [68Ga]3 was confirmed in blockade studies by co-injecting c RGDfK) shown in Fig. 2b, which resulted in a significant reduction of tumor accumulation (0.67 ± 0.10 %IA g− 1) (P value < 0.0001) and reduced uptake in all other organs.
We further confirmed the blockade of neuronal activity in the BoNT/E-treated cortex by evaluating Egr-1 expression (Fig. 2B).
Instead, we confirmed p38MAPK blockade by SB by means of ELISA.
We next confirmed the blockade of TGF-β signalling by EW-7197 in vivo.
The effect of PKA-Cα downregulation was also confirmed by blockade of forskolin-stimulated CREB phosphorylation in these cells (Fig. 7B, panel a, lane 4 versus lane 2).
Electrophysiological characterization of AGRP neurons showed the presence of SK-mediated tail currents, confirmed by blockade with the highly selective SK channel antagonist, apamin.
Moreover, we confirmed the blockade of Wnt-1 signaling in A-204 cells after Wnt-1 siRNA treatment (nonsilencing siRNA as control) by western blot analysis.
