MOESM2 of ICAM-1-related long non-coding RNA: promoter analysis and expression in human retinal endothelial cells

Additional file 2: Table S2. Predicted transcription factor binding sites within genome evolutionary rate profiling (GERP)-constrained elements

MOESM3 of ICAM-1-related long non-coding RNA: promoter analysis and expression in human retinal endothelial cells

Additional file 3: Table S3. Description of phenotype-nucleotide polymorphisms (SNPs) predicted to influence transcription of the human ICR gene

MOESM1 of ICAM-1-related long non-coding RNA: promoter analysis and expression in human retinal endothelial cells

Additional file 1: Table S1. Sources for eutherian genomes used in genome evolutionary rate profiling (GERP) and chromosomal locations of GERP-constrained elements

MOESM4 of ICAM-1-related long non-coding RNA: promoter analysis and expression in human retinal endothelial cells

Additional file 4: Table S4. Primer pairs and product sizes for gene transcripts studied in human retinal endothelial cells. References are provided for primers sequences sourced from the literature

<i>T. gondii</i> tachyzoites migrate through human retina.

<p>(A). Representative photomicrographs showing <i>T. gondii</i> tachyzoites within the retina 8 hours following addition of a suspension of 1.5×10⁷ live parasites to a human posterior eyecup. Tachyzoites are identified by immunostaining for the parasite SAG-1 antigen. Fast Red with hematoxylin counter stain. Original magnification: 1000X. Arrows indicate tachyzoites. Negative control sections showed no positive staining. (B). Graph showing number of tachyzoites counted within retinas from eyecups incubated with live or heat-killed tachyzoites. Columns  =  mean. Error bars  =  standard error of mean. (C). Pie chart showing mean percentage of tachyzoites located at different retinal layers. NFL  =  nerve fiber layer; GCL  =  ganglion cell layer; IPL  =  inner plexiform layer; INL  =  inner nuclear layer; OPL  =  outer nuclear layer; ONL  =  outer nuclear layer. Data shown in (B) and (C) were generated in experiments using 3 paired human cadaver posterior eyecups.</p

<i>T. gondii</i> tachyzoites infect human retinal glial cells in preference to neurons.

<p>(A). Immediately prior to infection, dissociated human retinal cultures presented a layer of glial cells with neurons positioned above. Original magnification: 100X. (B and C). Expression of (B) neuron specific enolase (NSE) (red), as detected by rabbit polyclonal anti-human NSE antibody and Alexa Fluor 594-conjugated donkey anti-rabbit immunoglobulin (Ig)G antibody and (C) glial fibrillary acidic protein (GFAP) (red), as detected by sheep polyclonal anti-human GFAP antibody and Alexa Fluor 594-conjugated donkey anti-sheep IgG antibody. <i>T. gondii</i> tachyzoites express YFP (green). Original magnification: 630X. Negative control cultures showed no positive staining. (D). Graph showing percentage growth of tachyzoites in Y79 human retinoblastoma cells and MIO-M1 human Müller glial cells, plus positive control human foreskin fibroblasts (FF), over a 24-hour period. n = 7–8 wells/condition. Columns  =  mean. Error bars  =  standard error of mean. Representative of two independent experiments.</p

Distribution of MSCs.

<p>A. phase contrast microphotograph of bone marrow derived mesenchymal stem cells at passage 2. B. MSCs were preincubated with PKH26 before intravenous injection. C. PKH26 labeled MSCs in the retina two weeks after intravenous injection (arrows); blood vessels were perfused with FITC-dextran (green). D–F. showing PKH26 labeled MSCs in the retinal section (D, arrows pointing PKH26 labeled MSCs; double arrows indicating background staining in debris zone); sections counterstained with DAPI (E); F. merged image from D&E showing PKH26 labeled MSCs counterstained with DAPI (scale bar equals 100 µm).</p

Upregulation of trophic factors.

<p>A. Semi-quantitative RT-PCR for CNTF, bFGF, BDNF and beta actin. Lane 1: RNA isolated from MSC prior to injection; Lane 2–4: RNA isolated from retinas treated with MSC; Lane 5–7: RNA isolated from non-treated control retinas. B. Densitometry analysis of CNTF, BDNF and bFGF in treated versus untreated samples. Beta actin was used to normalize the data for comparison. Level of CNTF and BDNF in the treated retinas were significantly higher than non-treated controls (p<0.05), while the level of bFGF in MSC treated retina did not increase significantly. C–J: confocal images of retinal sections double stained with antibodies to CNTF (green) and GFAP (red), counterstained with DAPI (blue in C and G) from MSC treated and controls. Strong CNTF staining in MSC treated retina (D) compared with untreated control (H); E&I: retinal sections stained with GFAP (red) showing upregulation of GFAP in Müller glia in both MSC treated and untreated control; F&J: merged images showing colocalization of CNTF and GFAP in MSC treated retina (F), which was not observed in untreated control (J) (Scale bar equals 50 µm).</p

Vascular protection.

<p>A–F: Retinal whole mount was stained with NADPH-diaphorase: A. typical vascular pathology in the eye at P90 in untreated RCS rat: vascular complexes (abnormal vessels associated with RPE cells) were mainly located around the optic nerve disc (arrows) and spread peripheral with age. B. vascular complexes in the middle to peripheral retina (arrows). C. high power image showing vascular complexes (arrows) from B. D. RCS retina treated with MSCs at P90: the vascular complexes were dramatically reduced around the optic nerve disc. E. two vascular complexes (arrows) in the middle field of the retina. F. high power image from E showing vascular complexes (arrow). G–L. animal was perfused with FITC-dextran, whole mount was prepared: G. typical vascular leakage, mainly around the optic disc in untreated eye at P90. H–K. high power images from G showing vascular leakage (arrows in H) and abnormal vessels (arrows in I–K). L. MSC treated retina, the vascular leakage around the optic nerve disc was greatly reduced. M&N. high power images from L showing much reduced leakage (arrows in M) and small abnormal vessels (arrow in N) (Scale bars equal 250 µm for A, D, G &L; 100 µm for F).</p

TEAD4 is upregulated and alternatively spliced within the eye in an animal model of ocular ischemic disease.

<p>RT-PCR for TEAD4 from choroidal, retinal and iris tissue isolated from a non-human primate eye (<i>Rhesus macaque</i>) 24 hrs after occlusion of the central retinal artery (CRAO), indicates that the full length TEAD4₄₃₄ transcript is increased and the TEAD4₁₄₈ enhancer isoform is produced in the lasered eye. <b>L</b> =  Lasered CRAO eye; <b>C</b>  =  control eye.</p