GLUT1 is up-regulated in SIRT6-KO retina.

<p>a) GLUTl immunoreactivity in cross-section of WT and SIRT6-KO mice retina. Ganglion Cell Layer (GCL), Inner Plexiform Layer (IPL), Inner nuclear Layer (INL) Outer Plexiform Layer (OPL), Outer Nuclear Layer (ONL), Retinal Pigment Epithelium (RPE). GLUT1 protein (b) and mRNA levels (c) were determined by Western blot and RT-PCR respectively. β-actin was used as loading control. Data are mean ± SE (n = 6 eyes/group) **p<0.01</p

SIRT6 is active in the mouse retina.

<p>a) H3K56 acetylation is shown by immunofluoescence. b) Representative Western blot showing protein levels of SIRT6 and the acetylation levels of H3K56 and H3K9 in chromatin preparations from WT and KO mice retinas. Total H3 was used for normalization. c) Quantification of the intensity of bands was determined by using the ImageJ and is represented as arbitrary units. Data are mean ± SE (n = 6 eyes/group). **p<0.01, ***p<0.001</p

Grm6 is down-regulated in SIRT6-KO retinas.

<p>Whole retina mRNA from WT and KO mice was used to profile the expression of several key genes of glutamate receptors involved in the synaptic transmission in an Affymetrix Mouse Gene 2.1 ST DNA microarray. a) Heatmap representing the hierarchical cluster analysis shows the differential expressed mRNAs between WT and SIRT6 KO retinas. The graphic depicts the expression levels of ionotropic AMPA glutamate receptors (Gria1–4), Glutamate receptor, ionotropic kainate (Grik1-2-4-5), Glutamate [NMDA] receptors (Grin1-2a-c) and metabotropic glutamate receptors (Grm1–8). The expression data for the hierarchical clustering image has been row normalized to a range of zero to one with blue representing the row minimum and red representing the row maximum. b) RNA was purified from SIRT6 WT and KO retinas, and Grm6 levels analyzed by RT-PCR. c) immunofluorescence was performed in SIRT6 WT and KO retinas with the indicated antibodies. PKC-alpha was used as a marker for ON bipolar cells. Ganglion Cell Layer (GCL), Inner Plexiform Layer (IPL), Inner nuclear Layer (INL), Outer Plexiform Layer (OPL), Outer Nuclear Layer (ONL), Retinal Pigment Epithelium (RPE). Data are mean ± SE (n = 4) **p<0.01 d) Representative fluorescent images of TUNEL analysis performed in WT and SIRT6 KO retinal sections. Apoptotic nuclei (bright green dots) labeled with fluorescein-dUTP were visualized by fluorescence microscopy. Data are mean ± SE (n  = 3) **p<0.01</p

Retinal functional evaluation.

<p>Representative scotopic (A) and photopic (C) electroretinograms from WT and SIRT6-KO mice at different light intensities (dBs). Plots B and D depict average amplitudes of <i>a</i>-wave and <i>b</i>-wave. Note that the fold decrease of the scotopic <i>a</i>-wave amplitude (8) is greater than the fold decrease of the photopic <i>a</i>-wave amplitude (2,5). Data are mean ± SE (n =  4). **p<0.01, ***p<0.001.</p

High-throughput gene expression profiling of memory differentiation in primary human T cells-3

Arker genes or control genes (grey symbols) between memory-phenotype and naive CD4 T cells. Values > 0 show relative increase in expression in memory-phenotype CD4 T cells; < 0 show relative increase in naive CD4 T cells. Expression levels for corresponding genes measured by LMA are plotted on Y axis; by Affymetrix U133A microarray on X-axis. Each point represents the mean values from cells sorted from ~4 – 6 subjects. Rand value refer to Spearman correlation coefficient.<p><b>Copyright information:</b></p><p>Taken from "High-throughput gene expression profiling of memory differentiation in primary human T cells"</p><p>http://www.biomedcentral.com/1471-2172/9/44</p><p>BMC Immunology 2008;9():44-44.</p><p>Published online 1 Aug 2008</p><p>PMCID:PMC2529265.</p><p></p

High-throughput gene expression profiling of memory differentiation in primary human T cells-5

Od T cells were calculated and the deviation of that data point from its corresponding mean were calculated. The fraction of data points in each of 12 bins of fold deviation values is shown, representing 1100 data points (two differentiation states × 55 transcripts × 6 replicates). (B) Cumulative Z-score for multiple replicates of memory-phenotype (purple) or naive (green) CD4 T cells measured by the method carried out using robotic automation. Values > 0 show relative increase in expression of memory-phenotype signature genes; < 0 show relative increase of naive signature genes.<p><b>Copyright information:</b></p><p>Taken from "High-throughput gene expression profiling of memory differentiation in primary human T cells"</p><p>http://www.biomedcentral.com/1471-2172/9/44</p><p>BMC Immunology 2008;9():44-44.</p><p>Published online 1 Aug 2008</p><p>PMCID:PMC2529265.</p><p></p