Journal: The Journal of Biological Chemistry

Article Title: TGF-β/SMAD3 Pathway Stimulates Sphingosine-1 Phosphate Receptor 3 Expression

doi: 10.1074/jbc.M116.740084

Figure Lengend Snippet: Up-regulation of S1PR3 in human lung adenocarcinomas. A, qPCR quantitation of S1PR3 mRNA in cDNA arrays of human lung adenocarcinoma specimens (OriGene, HLRT101 and HLRT105). **, p < 0.01, Student's t test. B, qPCR quantitation of S1PR2 mRNA in a cDNA array of human lung cancers (OriGene, HLRT105). **, p < 0.01, Student's t test. C, HEK293 cells were transfected with S1PR3 or pcDNA vector. Transfected cells were immunostained with anti-S1PR3 (Cayman Chemical) (IMF, left panels). Arrows, nonspecific fluorescent precipitates used for image orientation. Scale bar = 33 μm. D, anti-S1PR3 staining of human lung adenocarcinoma tumor microarray (Accumax 306). AdC, adenocarcinoma; N, adjacent normal lung tissue. E, immunostaining intensity was quantitated with the National Institutes of Health ImageJ software. Data, analyzed with GraphPad Prism 5 software, are shown as mean ± S.E. Statistical significance was analyzed by Student's t test. F, representative images of anti-S1PR3 staining of human lung adenocarcinoma and the respective adjacent normal lung epithelial tissue. G, quantitation of anti-S1PR3 staining of human lung squamous carcinoma microarray (Accumax 306). Data are mean ± S.E. Statistical significance was analyzed by Student's t test. H, representative images of anti-S1PR3 staining of human lung squamous carcinoma and the respective adjacent normal lung epithelial tissue.

Article Snippet: The sense and antisense primers used for qPCR analysis are: human and mouse S1PR1, sense, 5′-ATC ATG GGC TGG AAC TGC ATC A-3′, antisense, 5′-CGA GTC CTG ACC AAG GAG TAG AT-3; human and mouse S1PR2, sense, 5′-CAG ACG CTA GCC CTG CTC AAG A-3′, antisense, 5′-TAG TGG GCT TTG TAG AGG A-3′; human and mouse S1PR3, sense, 5′-ACA ACC GCA TGT ACT TTT TCA T-3′, antisense, 5′-TAC TGC CCT CCC TGA GGA ACC A-3′; human S1PR4, sense, 5′-GGG CCA TCT TCC GCC TGG TG-3′, antisense, 5′-TGC CCC GCA GGT ACT CCT GG-3′; human S1PR5, sense, 5′-GGC GCG CAC CTG TCC TGT AC-3′, antisense, 5′-TCG GGT CTC TGC CGC AGG AG-3′; human and mouse SphK1, sense, 5′-AAA CCC CTG TGT AGC CTC CC-3′, antisense, 5′-AGC AGG TTC ATG GGT GAC AG-3′; human SphK2, sense, 5′-GCA CAG CAA CAG TGA GCA-3′, antisense, 5′-GAG CCT GAG TGA GTG GGA-3′; porcine TGF-β, sense, 5′-GCA CGT GGA GCT ATA CCA GAA-3′, antisense, 5′-CAT CAA AGG ACA GCC ACT CC-3′; human GAPDH, sense, 5′-GAA GGT GAA GGT CGG AGT-3′, antisense, 5′-GAA GAT GGT GAT GGG TTT C-3′; and mouse GAPDH, sense, 5′-CAC CTT CGA TGC CGG GGC TG-3′, antisense, 5′-GGC CAT GAG GTC CAC CAC CC-3′. cDNA array analysis of mRNA levels of S1PR3 was performed using TissueScan qPCR arrays (HLRT101 and HLRT105, OriGene) following the manufacturer's instructions.

Techniques: Quantitation Assay, Transfection, Plasmid Preparation, Staining, Microarray, Immunostaining, Software

Journal: Journal of Cancer

Article Title: SKA2-mediated transcriptional downregulation of the key enzyme of CoQ 10 biosynthesis PDSS2 in lung cancer cells

doi: 10.7150/jca.79058

Figure Lengend Snippet: Functional implication of SKA2-PDSS2 axis in lung cancer. ( a ) Boxplots of PDSS2 expression in TCGA lung cancer dataset. ( b ) PDSS2 expression levels were analyzed with qRT-PCR using OriGene TissueScan lung cancer panels (HLRT103). The stages were divided into I, II, III, and IV. “N” means normal. The wide bars denote the mean levels of PDSS2, while the narrow ones represent standard deviation away from the mean. ( c ) Survival analysis in relation to SKA2 and PDSS2 expression in lung cancer. Kaplan-Meier plot of overall survival of lung cancer patients in the GSE13213 dataset, the p-values were calculated using Log-rank test. ( d ) PDSS2 abrogates SKA2-promoted lung cancer cells growth. Cells were transfected with indicated plasmids and cell growth was continuously monitored by JULI Stage Real-time Cell History Recorder. ( e, f ) Cell motility analysis. Cell motility was also captured every 15 minutes for 12 hours by the aforementioned instrument. Motile trajectory, mean total migration distances and mean migration speeds are presented.

Article Snippet: TissueScan lung cancer panels (HLRT103) containing prepared cDNAs were purchased from OriGene (Rockville, MD).

Techniques: Functional Assay, Expressing, Quantitative RT-PCR, Standard Deviation, Transfection, Migration