Phospho-NFAT1 (Ser326) Antibody - #AF8011

FIGURE 5 Aβ oligomer-induced expression levels of BACE1 and inactive NFAT1 by interference with Nav1.6, TTX, EGTA, and KB-R7943. Representative immunoblots (a) and densitometry analysis of P-NFAT1 (inactive) (b), NFAT1 (active) (c), BACE1 (d), and Nav1.6 (e) expression levels in SH-SY5Y cells after treatment with siNav1.6, 1 μM TTX, 0.5 mM EGTA, and 5 μM KB-R7943 under induced Aβ oligomers condition. (f) Relative mRNA expression of RT-qPCR showing the expression level of BACE1 in the SH-SY5Y cells after treatment with siNav1.6, 1 μM TTX, 0.5 mM EGTA, and 5 μM KB-R7943 under induced Aβ oligomers condition. (g) Intracellular calcium levels in SH-SY5Y cell after treatment with NC, siNav1.6, 1 μM TTX, 0.5 mM EGTA, and 5 μM KB-R7943 under induced Aβ oligomers condition. (h) SH-SY5Y cells were either treated with 1 μM TTX, 0.5 mM EGTA, and 5 μM KB-R7943 or transfected with Nav1.6 siRNA in the presence of Aβ oligomers and immunostained for NFAT1 and DAPI. Scale bar: 100 μm. (i) The ratio of nuclear NFAT1 to the cytoplasmic NFAT1. Here, P-NFAT1, BACE1, and Nav1.6 were normalized to γ-tubulin in the Western blot, whereas BACE1 was normalized to ACTIN in the RT-PCR. Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, **p < 0.001. Representative immunoblots (j) and densitometry analysis (k) of Nav1.6, P-NFAT1, and NFAT1 protein expression in the brain of mice (WT and APP/PS1 treated with siNav1.6 or NC), n = 5 mice/group. (l) The proposed flowchart cycle depicting interference of Nav1.6’s effect on the progression of AD. Interference of Nav1.6 can reduce the Aβ oligomers-dependent transcription of BACE1, which then relieves intracellular calcium overload by inhibiting sodium-calcium reverse exchange channel and leads to an increase in non-activated NFAT1 expression levels. The reduced transcription of BACE1, in turn, decreases Aβ production and slows the progression of AD