Journal: Experimental & Molecular Medicine

Article Title: Crosstalk between FLS and chondrocytes is regulated by HIF-2α-mediated cytokines in arthritis

doi: 10.1038/emm.2015.88

Figure Lengend Snippet: IL-6 and TNF-α have distinct effects on FLS and chondrocytes. ( a and b ) Mouse FLS ( a ) and chondrocytes ( b ) were infected with the indicated MOI of Ad- Epas1 , and the levels of secreted IL-6 and TNF-α in the culture media were determined by ELISA. ( c and d ) The indicated amounts of recombinant mouse IL-6 protein were used to treat FLS ( c ) or chondrocytes ( d ) for 24 h, and the mRNA levels of Mmp3 and Mmp13 were detected by qRT-PCR. ( e and f) Mouse FLS ( e ) and articular chondrocytes ( f ) were treated with the indicated doses of recombinant mouse TNF-α protein for 24 h, and the levels of the indicated catabolic factors were determined by qRT-PCR. Values are presented as means±s.e.m. (* P <0.01, ** P <0.005).

Article Snippet: For isolation of chondrocytes, mouse cartilage tissues were isolated from the femoral condyles and tibial plateaus of wild-type and Il6 −/− mice, and chondrocytes were extracted by digestion with 0.2% collagenase type II, as described previously., Chondrocytes or FLS were treated with the indicated amounts of recombinant mouse IL-6 protein (Merck-Millipore, Billerica, MA, USA), recombinant mouse TNF-α protein (Merck-Millipore) or 800 MOI (multiplicity of infection) of empty adenovirus (Ad-C) or HIF-2α-expressing adenovirus (Ad- Epas1 ) for 24 h with or without subsequent treatment with the anti-TNF-α blocking antibody (R&D Systems Inc., Minneapolis, MN, USA).

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Recombinant, Quantitative RT-PCR

Journal: Experimental & Molecular Medicine

Article Title: Crosstalk between FLS and chondrocytes is regulated by HIF-2α-mediated cytokines in arthritis

doi: 10.1038/emm.2015.88

Figure Lengend Snippet: Blockade of TNF-α secretion in co-cultured FLS or chondrocytes. Mouse chondrocytes or FLS were infected with or without 800 MOI of Ad-C or Ad- Epas1 and incubated with FLS ( a ) or chondrocytes ( b ), respectively. After 12 h, the indicated amounts (μg) of TNF-α-neutralizing antibody were added. After an additional 12 h, the transcripts of the indicated catabolic factors were detected by RT-PCR and qRT-PCR. Values are presented as means±s.e.m. (* P <0.05, ** P <0.005).

Article Snippet: For isolation of chondrocytes, mouse cartilage tissues were isolated from the femoral condyles and tibial plateaus of wild-type and Il6 −/− mice, and chondrocytes were extracted by digestion with 0.2% collagenase type II, as described previously., Chondrocytes or FLS were treated with the indicated amounts of recombinant mouse IL-6 protein (Merck-Millipore, Billerica, MA, USA), recombinant mouse TNF-α protein (Merck-Millipore) or 800 MOI (multiplicity of infection) of empty adenovirus (Ad-C) or HIF-2α-expressing adenovirus (Ad- Epas1 ) for 24 h with or without subsequent treatment with the anti-TNF-α blocking antibody (R&D Systems Inc., Minneapolis, MN, USA).

Techniques: Cell Culture, Infection, Incubation, Reverse Transcription Polymerase Chain Reaction, Quantitative RT-PCR

Journal: Experimental & Molecular Medicine

Article Title: Crosstalk between FLS and chondrocytes is regulated by HIF-2α-mediated cytokines in arthritis

doi: 10.1038/emm.2015.88

Figure Lengend Snippet: The receptors for IL-6 and TNF-α show distinct expression patterns in HIF-2α-overexpressing FLS and chondrocytes. ( a and b ) Mouse FLS or chondrocytes were infected with Ad- Epas1 or empty virus at an MOI of 800 for 24 h, and microarray analyses were performed on chondrocytes and FLS. Microarray data for IL-6 receptors ( Il6ra and Il6st ) and TNF-α receptors ( Tnfrsf1a and Tnfrsf1b ) in Ad- Epas1 -infected FLS ( a ) and chondrocytes ( b ) are shown. Transcript expression levels are shown relative to those in Ad-C-treated cells. ( c) Schematic diagram summarizing the regulatory mechanism governing the in vitro interaction between FLS and chondrocytes in the presence of HIF-2α overexpression.

Article Snippet: For isolation of chondrocytes, mouse cartilage tissues were isolated from the femoral condyles and tibial plateaus of wild-type and Il6 −/− mice, and chondrocytes were extracted by digestion with 0.2% collagenase type II, as described previously., Chondrocytes or FLS were treated with the indicated amounts of recombinant mouse IL-6 protein (Merck-Millipore, Billerica, MA, USA), recombinant mouse TNF-α protein (Merck-Millipore) or 800 MOI (multiplicity of infection) of empty adenovirus (Ad-C) or HIF-2α-expressing adenovirus (Ad- Epas1 ) for 24 h with or without subsequent treatment with the anti-TNF-α blocking antibody (R&D Systems Inc., Minneapolis, MN, USA).

Techniques: Expressing, Infection, Virus, Microarray, In Vitro, Over Expression

Journal: Malaria Journal

Article Title: Differential gene expression mediated by 15-hydroxyeicosatetraenoic acid in LPS-stimulated RAW 264.7 cells

doi: 10.1186/1475-2875-8-195

Figure Lengend Snippet: Select Gene Expression Changes Mediated by 15-HETE a

Article Snippet: Following RT, all assays were performed with Applied Biosystems TaqMan FAM labeled 20× probes: Arf3 (Taqman assay Mm00500194_m1), Cldn11 (Mm00500915_m1), Cxcl11 (Mm00444662_m1), Mapk14 (Mm00442497_m1), Prdx1 (Mm01621996_s1), Sdc1 (Mm00448918_m1), and Egr1 (Mm00656724_m1).

Techniques: Gene Expression, Binding Assay, Ubiquitin Proteomics, Virus, Transduction, Membrane, Activation Assay

Journal: Malaria Journal

Article Title: Differential gene expression mediated by 15-hydroxyeicosatetraenoic acid in LPS-stimulated RAW 264.7 cells

doi: 10.1186/1475-2875-8-195

Figure Lengend Snippet: Quantitative real-time RT-PCR validation of microarray results . RAW 264.7 cells were stimulated with 0.1 μg/mL LPS and treated with 40 μM 15-HETE for 24 h prior to RNA extraction. Fold-changes (treated stimulated cells relative to stimulated cells) are shown ( ± 99% confidence interval for quadruplicate measurements of n = 3 biological replicates). Abbreviations: Arf3 (ADP-ribosylation factor 3), Cldn11 (claudin 11), Cxcl11 (chemokine (C-X-C motif) ligand 11), Egr1 (early growth response 1), Mapk14 (mitogen-activated protein kinase 14), Prdx1 (peroxiredoxin 1), and Sdc1 (syndecan 1).

Article Snippet: Following RT, all assays were performed with Applied Biosystems TaqMan FAM labeled 20× probes: Arf3 (Taqman assay Mm00500194_m1), Cldn11 (Mm00500915_m1), Cxcl11 (Mm00444662_m1), Mapk14 (Mm00442497_m1), Prdx1 (Mm01621996_s1), Sdc1 (Mm00448918_m1), and Egr1 (Mm00656724_m1).

Techniques: Quantitative RT-PCR, Biomarker Discovery, Microarray, RNA Extraction

Journal: Human reproduction (Oxford, England)

Article Title: Identification of new ovulation-related genes in humans by comparing the transcriptome of granulosa cells before and after ovulation triggering in the same controlled ovarian stimulation cycle.

doi: 10.1093/humrep/deu008

Figure Lengend Snippet: Figure 2 Validation of microarray results by qRT-PCR in 11 selected genes in 7 paired samples of GC prior to and 36 h after rhCG triggering. All validated genes were significantly differentially expressed before compared with after rhCG in the qRT-PCR experiment (P-values, ratio paired t-test): CD24: P ¼ 0.0069, AREG: P ¼ 0.0015, PTGS2: P ¼ 0.0004, COL6A3: P ¼ 0.0007, HSD11B1: P ¼ 0.0001, HSD11B2: P ¼ 0.0132, INHBA: P ¼ 0.0012, CCNA2: P ¼ 0.0037, PTTG1: P ¼ 0.0049, IGF2: P ¼ 0.0064 and CYP19A1: P ¼ 0.0010. GC, granu- losa cells.

Article Snippet: Validation of differential expression by quantitative reverse transcriptase PCR (qRT-PCR) The following genes were testedby pre-designed TaqManw Gene Expression Assays (gene symbol/catalogue number): CD24 (Hs03044178_g1), AREG (Hs00950669_m1), HSD11B1 (Hs01547870_m1), HSD11B2 (Hs003886 69_m1), PTGS2 (Hs00153133_m1), COL6A3 (Hs00915125_m1), CYP19A1 (Hs00903413_m1), IGF2 (Hs04188276_m1), PTTG1 (Hs00851754_u1), CCNA2 (Hs00996788_m1) and INHBA (Hs01081598_m1) as well as the Endogenous Control Assays for human glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (Applied Biosystems/Life Technologies Europe, Nærum, Denmark).

Techniques: Biomarker Discovery, Microarray, Quantitative RT-PCR

Journal: Evolution & development

Article Title: Abundant genetic variation in transcript level during early Drosophila development

doi: 10.1111/j.1525-142X.2008.00281.x

Figure Lengend Snippet: Maternal and zygotic gene expression

Article Snippet: Found on Affymetrix Microarray Expressed Varying among lines Varying over time Increasing Declining Changes during development 2089 (AST7 * ) 2065 ** (ST4) 1215 (ST5) 950 (ST6) 698 (ST7) 224 (ST8) 491 (ST9) Strictly maternal 27 (AST12) 27 (ST10) 21 (ST11) 21 (ST12) 19 (ST13) 0 19 (ST14) Maternal 49 (AST13) 50 (ST15) 41 (ST16) 35 (ST17) 22 (ST18) 2 (ST19) 20 (ST20) Strictly Zygotic 53 (AST19) 57 (ST27) 29 (ST28) 25 (ST29) 22 (ST30) 19 (ST31) 3 (ST32) Zygotic 532 (AST18) 514 (ST21) 176 (ST22) 141 (ST23) 127 (ST24) 89 (ST25) 38 (ST26) Open in a separate window * AST stands for Supplementary Table in Arbeitman et al. 2002 ; and ST is for the Supplementary Table in this article.

Techniques: Microarray