Representative immunohistochemical staining of GFAP in SNpc.

<p><b>A</b> and <b>B</b>, immuno-staining against GFAP antibody. <b>A'</b> and <b>B'</b>, immuno-fluorescent staining against TH antibody. <b>A''</b> and <b>B''</b>, merging of immuno-stainings against GFAP and TH antibodies, the later was represented with a pseudo-color. SNpc was highlighted in each panel. Inset, a higher magnification of SNpc for each picture. Scale bar, 200 µm.</p

Validation of microarray results in SNpc by real time quantitative PCR.

<p>Relative log2 fold changes of mRNA were presented comparing late middle-aged (18months old) to young (2month old) mice. Positive numbers corresponded to up-regulated genes, whereas negative numbers indicated down-regulated genes. *, <i>p</i><0.05 comparing 18 to 2 months old mice using Student’s <i>t</i>-test. <i>n</i> = 4 replicates for each age group.</p

Gene ontology analysis of age-dependent genes in SNpc and VTA comparing 18 to 2 months old mice.

<p>Gene ontology analysis of age-dependent genes in SNpc and VTA comparing 18 to 2 months old mice.</p

Volcano plots illustrating differential gene expression in SNpc (A) & VTA (B) comparing late middle-aged (18months old) to young (2month old) mice.

<p>Differentially expressed genes were distributed in the top left and top right sections of each figure, corresponding <i>pFDR</i> <0.05 and fold change<−1.3 and <i>pFDR</i> <0.05 and fold change >1.3, respectively.</p

Age-associated genes specific to mouse SNpc.

*<p>Comparing 18 months old to 2 months old mice, positive numbers corresponded to up-regulated genes, whereas negative numbers indicated down-regulated genes.</p

Top 15 biological functions over-represented in aged (≥ 24 months old) mouse brain in comparison to SNpc and VTA in late middle-aged (18 months old) mice.

<p><b>A</b>. VTA; <b>B</b>. Top 120 probe sets with the lowest <i>pFDR</i> values in VTA. Threshold: <i>p = </i>0.05.</p

Gene ontology analysis of age-associated genes specific to SNpc.

<p>Gene ontology analysis of age-associated genes specific to SNpc.</p

SNpc and VTA dissection from mouse brain.

<p>Brain sections were labeled against tyrosine hydroxylase antibody without (<b>A</b>) and after (<b>B</b>) SNpc dissection, and without (<b>C</b>) and after (<b>D</b>) VTA dissection. Scale bar: 500 µm. <b>E</b>. Relative <i>Calb1</i> mRNA level. *, <i>p</i><0.05 comparing VTA to SNpc (<i>n</i> = 4 in each group).</p

Expression profile of miR-200f and EMT-transcriptional inducers in breast tumors.

<p>The expression levels of miR-200 family members (<b>A</b>) and EMT-transcriptional inducers (<b>B</b>) were quantified by qRT-PCR in 70 breast cancer samples. Data are depicted as box-and-whisker plots. Adjusted <i>p</i>-values are shown where significant differences were found (Wilcoxon rank-sum test). ER+ (estrogen receptor positive tumors), HER2+ (HER2-positive tumors), TN (triple negative; ER−, PR−, HER2−), MBC (metaplastic breast carcinomas). (<b>C</b>) Upper heatmap represents Pearson coefficient (R) correlation values between the expression of EMT-transcriptional inducers and miR200f members. Bottom heatmap depicts the level of statistical significance (P) of the correlations.</p

miR-200f and EMT-transcriptional inducers are modulated during <i>in vitro</i> EMT in breast basal cell lines.

<p>Expression of (<b>A</b>) miR-200f and (<b>B</b>) EMT-transcriptional inducers was evaluated by qRT-PCR in the sorted epithelial (EpCAM+) and mesenchymal (Fibros) subpopulations within MCF12A and Myo1089 cell lines. Data are normalized to the expression of <i>RNU48</i> and <i>18S</i> respectively. Bars represent mean expression changes ±SE in Fibros subpopulation relative to EpCAM+ cells (baseline). Three biological replicates were measured. Moderated t-test was performed to evaluate statistical significance (<i>p</i><0.001 for all miRNAs analyzed in panel A. <i>NS</i>, non significant). (<b>C</b>) Methylation status of <i>miR-200f</i> promoter sequences in EpCAM+ and Fibros subpopulations within MCF12A and Myo1089 cell lines. Histogram bars represent averaged methylation levels (±SE) for the promoter sequences of <i>miR-200f loci</i>. Statistically significant differences are indicated (*<i>p</i><0.001, Student’s t-test).</p