Increased PDIA3 protects from METH toxicity.

<p>Examination of SK-N-BE(2) cells for expression and knockdown for PDIA3 by (A) western blot and (B) immunofluorescence. Scale bar = 10 µm. (C) Increased METH cytotoxicity in PDIA3 knockdown cells compared to the PDIA3 expressors. Cytotoxicity was assessed by a lactate dehyrogenase assay following 48 hrs exposure to 500 µM METH. ***p<0.001, unpaired Student's t-test.</p

Summary of microarray and qRT-PCR analyses comparing differences in gene expression between stable BE(2)M17 clones expressing miR-142 and miR-null.

<p>Genes that were found to be down-regulated in the miR-142 clones with a p-value <0.001 in the microarray analysis are shown here. Only MAOA and AKAP12 showed significant (p < 0.05) down-regulation in the post-validation experiment using RT-PCR.</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

PDIA3 is increased by METH in rodents <i>in vivo</i>.

<p>Time course of gene expression, determined by qRTPCR, in striatum from mice treated with 10 mg/kg METH at the indicated time points. (A) PDIA3 (B) HSPA5 (as a positive control). *p<0.05 as determined by a one-way ANOVA followed by a post hoc Tukey's test.</p

METH induction of PDIA3 <i>in vitro</i> in rodent neurons.

<p>Primary rat striatal neurons (8 DIV) stained for levels and distribution of PDIA3 after 250 µM METH treatment for 24 h along with MAP2 staining for neuronal structure and DAPI for cell nucleus. Control cells show basal levels of PDIA3 while cells treated with METH show an increase and redistribution of PDIA3 along the neuronal processes indicated by arrowheads. Scale bar = 10 µm.</p

Increased PDIA3 suppresses ROS production.

<p>Cells treated with 500 µM METH for 1 h were assessed for production of ROS using DCFH-DA assay. A significant increase in ROS production is seen in cells knocked down for PDIA3, compared to PDIA3 expressors, both with and without METH treatment. Two-way ANOVA p<0.0001, Bonferroni post-tests. ***p<0.001. U – No treatment; M – METH treatment.</p

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

PDIA3 is increased in by METH in monkeys <i>in vivo</i>.

<p>Levels of PDIA3 mRNA expression from qRTPCR in brain regions from the two groups of animals, differing only by METH treatment. (A) caudate and (B) hippocampus. **p<0.01, unpaired Student's t-test.</p

List of genes up-regulated in both the caudate and hippocampus from SIV infected, METH and PBS administered monkeys.

<p>The gene symbol, gene name, p-value (unpaired Student's t-test) and fold change between the groups is indicated. Genes with p<0.01 and a fold change of >2 in both regions are shown.</p

miR-142 downregulates MAOA protein expression in human neurons.

<p>Human neurons grown in vitro for 11 days were transduced with either miR-142 or GFP lentivirus. On day 6 after transduction, cells were harvested and Western blot analysis was performed for MAOA. Representative Western blots from three human neuron donors indicating a decrease in MAOA protein level after miR-142 transduction compared to control GFP transduction are shown. Quantification of Western blots from 5 human neuron donors revealed a significant decrease in MAOA level after miR-142 transduction (fold change -1.9, p<.05). Blots were normalized to β-actin and paired t-test was performed.</p