Additional file 2: Figure S2. of Functional analysis of fatty acid binding protein 7 and its effect on fatty acid of renal cell carcinoma cell lines

Proliferation of TUHR14TKB cells transfected with an FABP7 expression vector. The proliferation of cells transfected with the FABP7 expression vector or lacZ expression vector was determined using an MTS assay. The data represent of five experiments. Transfectants were cultured in RPMI 1640 medium containing 5 mg/L blasticidin S HCl, 0.3 g/L G418, and 1 mg/L doxycycline hyclate with 1% FBS (a-b) or 10% FBS (c-d). a, c, TUHR14TKB lacZ. b, d, TUHR14TKB FABP7. (TIFF 1521 kb

Chlamydia pneumoniae Infection Increases Adherence of Mouse Macrophages to Mouse Endothelial Cells In Vitro and to Aortas Ex Vivo▿

Interactions between monocytes/macrophages and endothelial cells play an important role in the pathogenesis of atherosclerosis, and the adherence of monocytes to the arterial endothelium is one of the early events in atherogenesis. In the present study, peritoneal macrophages harvested from green fluorescent protein (GFP) transgenic mice were used to analyze how Chlamydia pneumoniae infection affects the adherence of GFP-macrophages to mouse endothelial cells in vitro and to the aorta from normolipidemic and hyperlipidemic mice ex vivo. In vitro studies showed that C. pneumoniae-infected GFP-macrophages adhered better than uninfected macrophages to endothelial cells and GFP-macrophages adhered better to infected than uninfected endothelial cells. The ex vivo studies showed that C. pneumoniae-infected macrophages adhered better than uninfected macrophages to aortas from both normolipidemic and hyperlipidemic C57BL/6J mice and apolipoprotein E (ApoE)-deficient mice. In contrast, adherence of C. pneumoniae-infected macrophages to the aortas of intercellular adhesion molecule 1 (ICAM-1) knockout mice was not enhanced, suggesting that ICAM-1 is crucial for activation of the adherence of C. pneumoniae-infected macrophages to the endothelium. In conclusion, the present study defined a homing mechanism by which C. pneumoniae promotes the adherence of mononuclear phagocytes to the endothelium at the site of atherosclerotic lesion formation to promote the progression of atherosclerosis

Analysis of the regulation of fatty acid binding protein 7 expression in human renal carcinoma cell lines

<p>Abstract</p> <p>Background</p> <p>Improving the treatment of renal cell carcinoma (RCC) will depend on the development of better biomarkers for predicting disease progression and aiding the design of appropriate therapies. One such marker may be fatty acid binding protein 7 (FABP7), also known as B-FABP and BLBP, which is expressed normally in radial glial cells of the developing central nervous system and cells of the mammary gland. Melanomas, glioblastomas, and several types of carcinomas, including RCC, overexpress FABP7. The abundant expression of FABP7 in primary RCCs compared to certain RCC-derived cell lines may allow the definition of the molecular components of FABP7's regulatory system.</p> <p>Results</p> <p>We determined <it>FABP7 </it>mRNA levels in six RCC cell lines. Two were highly expressed, whereas the other and the embryonic kidney cell line (HEK293) were weakly expressed <it>FABP7 </it>transcripts. Western blot analysis of the cell lines detected strong FABP7 expression only in one RCC cell line. Promoter activity in the RCC cell lines was 3- to 21-fold higher than that of HEK293. Deletion analysis demonstrated that three <it>FABP7 </it>promoter regions contributed to upregulated expression in RCC cell lines, but not in the HEK293 cell. Competition analysis of gel shifts indicated that OCT1, OCT6, and nuclear factor I (NFI) bound to the <it>FABP7 </it>promoter region. Supershift experiments indicated that BRN2 (POU3F2) and NFI bound to the <it>FABP7 </it>promoter region as well. There was an inverse correlation between <it>FABP7 </it>promoter activity and <it>BRN2 </it>mRNA expression. The FABP7-positive cell line's NFI-DNA complex migrated faster than in other cell lines. Levels of <it>NFIA </it>mRNA were higher in the HEK293 cell line than in any of the six RCC cell lines. In contrast, <it>NFIC </it>mRNA expression was lower in the HEK293 cell line than in the six RCC cell lines.</p> <p>Conclusions</p> <p>Three putative <it>FABP7 </it>promoter regions drive reporter gene expression in RCC cell lines, but not in the HEK293 cell line. BRN2 and NFI may be key factors regulating the expression of FABP7 in certain RCC-derived cell lines.</p

Complete genomic DNA sequence of the east asian spotted fever disease agent rickettsia japonica

Rickettsia japonica is an obligate intracellular alphaproteobacteria that causes tick-borne Japanese spotted fever, which has spread throughout East Asia. We determined the complete genomic DNA sequence of R. japonica type strain YH (VR-1363), which consists of 1,283,087 base pairs (bp) and 971 protein-coding genes. Comparison of the genomic DNA sequence of R. japonica with other rickettsiae in the public databases showed that 2 regions (4,323 and 216 bp) were conserved in a very narrow range of Rickettsia species, and the shorter one was inserted in, and disrupted, a preexisting open reading frame (ORF). While it is unknown how the DNA sequences were acquired in R. japonica genomes, it may be a useful signature for the diagnosis of Rickettsia species. Instead of the species-specific inserted DNA sequences, rickettsial genomes contain Rickettsia-specific palindromic elements (RPEs), which are also capable of locating in preexisting ORFs. Precise alignments of protein and DNA sequences involving RPEs showed that when a gene contains an inserted DNA sequence, each rickettsial ortholog carried an inserted DNA sequence at the same locus. The sequence, ATGAC, was shown to be highly frequent and thus characteristic in certain RPEs (RPE-4, RPE-6, and RPE-7). This finding implies that RPE-4, RPE-6, and RPE-7 were derived from a common inserted DNA sequence

Circular exhibition of the <i>Rickettsia japonica</i> YH genome.

<p>The outermost scale is marked for nucleic acid position in Mbp, and Region-1 and -2 (red). From the outside (track 1), gene positions and directions (clockwise on the outside and anti-clockwise on the inside) of each gene were classified and colored based on COGs <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071861#pone.0071861-Tatusov2" target="_blank">[45]</a>. Track 2 and 3: tRNA (red) and rRNA (blue), respectively. Track 4: GC skew, outside yellow and inside purple indicate values >0 and values <0 as calculated by (G−C/G+C) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071861#pone.0071861-Grigoriev1" target="_blank">[46]</a>. Track 5: innermost, GC contents. Based on the cumulus of the GC skew values, accompanied by other <i>Richettsia</i> genomes, a hypothetical origin, shown as <i>ori</i>, was determined and the base numbers were counted from the origin.</p

Multiple alignments of amino acid and DNA sequences, including inserted DNA sequences.

<p>Each panel shows A–C) DNA polymerase III alpha chain; D–F) tRNA ribosyltransferase-isomerase; G–I) phenylalanyl-tRNA synthetase subunit beta; A, D, G) amino acid alignments; B, E, H) structure models; C, F, I) DNA alignments. Letters in alignments indicate green, typical RPE-4s; blue, typical RPE-6s; purple, typical RPE-7s; red, extended RPE-4 regions; black, boundary regions of genes with inserted DNA sequences. Amino acids encoded by inserted DNA sequences were shown by red balls in the protein structures. Gene sequences were extracted from each genome sequence; <i>Rickettsia japonica</i> (this work); <i>R. conorii</i>, AE006914 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071861#pone.0071861-Ogata1" target="_blank">[13]</a>; <i>R. felis</i>, CP000053 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071861#pone.0071861-Ogata5" target="_blank">[28]</a>; <i>R. prowazekii</i>, AJ235269 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071861#pone.0071861-Andersson1" target="_blank">[29]</a>; <i>R. typhi</i>, AE017197 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071861#pone.0071861-McLeod1" target="_blank">[30]</a>; <i>Acetobacter pasteurianus</i> IFO 3283-01, AP011121 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071861#pone.0071861-Azuma2" target="_blank">[47]</a>. Protein Data Bank Identifiers (PDB IDs) for amino acid sequences of <i>Escherichia coli</i> DNA polymerase III alpha chain, <i>Bacillus subtilis</i> tRNA ribosyltransferase-isomerase, and <i>Thermus thermophilus</i> phenylalanyl-tRNA synthetase subunit beta are 2hnhA, 1yy3A, and 1b70B, respectively. J) The highly conserved short sequence in e-RPEs were illustrated using a sequence LOGO format <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071861#pone.0071861-Crooks1" target="_blank">[33]</a>.</p