Primary rat striatal neurons stained for levels and distribution of ATP1A3 after 250 µM METH treatment for 24 h along with MAP2 staining for neuronal structure.

<p>(A) and with synaptic marker synaptophysin (B). DAPI was used to stain cell nucleus. Cells treated with METH show an increase and redistribution of ATP1A3 along the neuronal processes indicated by arrowheads and in the nerve terminals (inset). Scale bar = 10 µm.</p

Synaptic proteins differentially regulated between the two groups of monkeys.

<p>Unpaired student t-test was used to determine the significance.</p

Western blot analysis on rat striatal neurons pre-treated with the ERK inhibitor U1026 followed by cytokines treatment (6 h) in absence (A) and presence (B) of METH treatment for 15 min show abrogation of the increase in ATP1A3 expression. Data represented as Mean ± SEM of three independent experiments.

<p>*p<0.05, **p<0.01 versus control, #p<0.05 versus cytokine ± METH treatment.</p

Western blot analysis on rat striatal neurons pre-treated with indicated cytokines (10 ng each) for 6 h followed by METH treatment for 24 h showing increased ATP1A3 expression compared to METH only and cytokine only treatments.

<p>(M – METH, I – Interferon-γ, T – TNF-α). Bar graphs showing significant increase in ATP1A3 expression post treatment with the cytokines. Data represented as Mean ± SEM of three independent experiments. *p<0.05, **p<0.01, ***p<0.001 versus METH treatment.</p

Western blot analysis on rat striatal neurons pre-treated with ERK1/2 JNK and p38 inhibitors U1026, SP600125, and SB203580 respectively followed by METH treatment shows U1026 blocks the METH-induced increase in ATP1A3 expression.

<p>(A) but SP600125 or SB203580 do not (B, C). Data represented as Mean ± SEM of three independent experiments. **p<0.01 versus control, ##p<0.01 versus METH treatment.</p

MTT assay showing viability of cells post treatment with varying amounts of METH for 24 h.

<p>**p<0.01, ***p<0.001 (A). (B) Representative western blot confirming up regulation of ATP1A3 in rat striatal neurons treated with indicated concentrations of METH for 24 h as compared to controls. (C) Bar graphs showing significant increase in ATP1A3 expression post METH treatment versus control. Data represented as Mean ± SEM of three independent experiments. *p<0.05.</p