Journal: Genes, brain, and behavior

Article Title: Functional conservation of MBD proteins: MeCP2 and Drosophila MBD proteins alter sleep

doi: 10.1111/gbb.12314

Figure Lengend Snippet: (a-a″) hMeCP2 expression (red) in OA neurons from an adult tdc2-gal4/UAS-mCD8:gfp; UAS-hMeCP2/+ male (anti-GFP, green; mAb nc82, labels neuropil regions, blue). (b–h) Sleep profiles of individual adult males averaged over 8 days from control and experimental groups. Controls: tdc2-gal4/+ (white), UAS-hMeCP2/+ (light grey), tdc2-gal4/+; UAS-dsRed/+ (dark grey) and experimental: tdc2-gal4/+; UAS-hMeCP2/+ (red). (b) Total sleep per 24-hr day is reduced in experimental males as compared to controls (Padj=0.0013; one-way ANOVA with Holm-Sidak’s multiple comparison test). (c) Eduction graph displaying 30 minute bins of averaged sleep (daytime/light phase: white bar; nighttime/dark phase: black bar, shaded grey). tdc2-gal4/+; UAS-hMeCP2/+ males displayed a reduction in the average amount of sleep during both day and night (arrows) as compared to controls. These deficits are quantified in (d) for Zeitgeber hours ZT04-10, (P<0.0001; two-tailed Mann Whitney test) and ZT14.5-22, (P<0.0001; two-tailed Mann Whitney test). (e–g) Sleep fragmentation in males expressing MeCP2 expression in OA neurons. As compared to controls, the average number of sleep bouts per day (e) is increased (Padj<0.0001) and weighted average bout length measured by the consolidation index (f) is reduced significantly in experimental males (Padj<0.0001). (g) The empirical cumulative distribution function (ECDF) demonstrating experimental males exhibit a greater proportion of short sleep bouts as compared to controls. (h) Latency to initiate sleep (the delay in minutes from the lights OFF to the time to the first sleep bout) is significantly reduced in tdc2-gal4/+; UAS-hMeCP2/+ males as compared to controls (Padj=0.0009; one-way ANOVA with Holm-Sidak’s multiple comparison test). Data are shown as means ± standard error of the mean (SEM).

Article Snippet: The empirical cumulative distribution (CDF) for sleep bouts was plotted using the ecdf function in MATLAB (The MathWorks, Natick, MA).

Techniques: Expressing, Control, Comparison, Two Tailed Test, MANN-WHITNEY

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: A novel miR-24-TCF1 axis in modulating effector T cell responses

doi: 10.4049/jimmunol.1601404

Figure Lengend Snippet: (A) Cumulative distribution frequency (CDF) plots depicting the effect of overexpression of miR-24 on mRNA expression of Th1 and Th17 cell-associated genes. Levels of mRNAs of Th1 or Th17 cell-associated genes bearing HITS-CLIP identified miR-24 sites (red line) were compared with mRNAs of all Th1 or Th17 cell-associated genes (black line). (B) HITS-CLIP analysis of putative miR-24 binding sites in the 3′UTR of TCF1. Reads from the 12 replicates have been stacked. Coordinates along the x-axis indicate nucleotide position relative to the beginning of the 3′UTR. The y axis indicates normalized read counts, which are square root transformed after individual library normalization (8). (C) Sequence alignment of putative miR-24 binding sites in the 3′UTR of TCF1. Mutations of the two corresponding miR-24 target sites are shown below. (D) Ratios of repressed luciferase activity of cells in the presence of WT or different mutated TCF1 3′ UTRs (M1: mutant 1, M2: mutant 2 or DM: double mutants) transfected with miR-24 compared with cells transfected with empty vector. (E) TCF1 mRNA expression results from the naive CD4+ T cell RNA-sequencing study was presented as a heat map. (F) qPCR of TCF1 mRNA levels, (G) FACS analysis and (H) mean fluorescence intensity (MFI) of TCF1 protein amounts in naïve CD4+ T cells from R24Tg mice or WT littermates were shown. (I) Expression of different TCF1 isoforms in naïve CD4+ T cells isolated from R24Tg or control mice were assessed by immunoblotting. Densitometric values of different TCF1 isoform were normalized to β-actin expression values and n-fold increase on the basis of each corresponding WT. All data are representative of at least 3 independent experiments (n = 6–7). *p<0.05, **p<0.01.

Article Snippet: Empirical cumulative distributions were computed using Matlab (R2014b) to display the log2(miRNA Tg/WT) against the cumulative frequency of Th1- or Th17- associated or all genes ( 13 ).

Techniques: Over Expression, Expressing, Binding Assay, Transformation Assay, Sequencing, Luciferase, Activity Assay, Mutagenesis, Transfection, Plasmid Preparation, RNA Sequencing, Fluorescence, Isolation, Control, Western Blot