Journal: Biosensors & bioelectronics

Article Title: Microarray method to monitor 40 intestinal bacterial species in the study of azo dye reduction

doi: 10.1016/j.bios.2004.04.011

Figure Lengend Snippet: Bacteria and the probe numbers in the microarray

Article Snippet: Anaerobic bacteria were cultured at 35 °C in either prereduced anaerobically sterilized (PRAS) Brain Heart Infusion (BIH) broth supplemented with vitamin K and hemin (Remel, Lenexa, KS, USA), inoculated under an oxygen-free cannula using 85% nitrogen, 10% hydrogen and 5% carbon dioxide, or on PRAS brucella blood agar plates supplemented with vitamin K and hemin (Remel). table ft1 table-wrap mode="anchored" t5 caption a7 Number Bacterial species and strain Probe number 1 B. thetaiotaomicron ATCC 29148 1, 2, 3 2 B. vulgatus ATCC 8482 4, 5, 6 3 B. fragilis ATCC 23745 7, 8, 9 4 B. distasonis ATCC 8503 10, 11, 12 5 C. clostridioforme ATCC 29084 13, 14, 15 6 C. leptum ATCC 29065 16, 17, 18 7 F. prausnitzii ATCC 27768 19, 20, 21 8 P. productus ATCC 27340 22, 23, 24 9 R. obeum ATCC 29174 25, 26, 27 10 R. bromii ATCC 27255 28, 29, 30 11 R. callidus ATCC 27760 31, 32, 33 12 R. albus ATCC 27210 34, 35, 36 13 B. longum ATCC 15707 37, 38, 39 14 B. adolescentis ATCC 15703 40, 41, 42 15 B. infantis ATCC 15697 43, 44, 45 16 E. biforme ATCC 27806 46, 47, 48 17 E. aerofaciens ATCC 25986 49, 50, 51 18 L. acidophilus ATCC 4356 52, 53, 54 19 E. coli ATCC 25922 55, 56, 57 20 E. faecium ATCC 19434 58, 59, 60 21 B. uniformis ATCC 8492 61, 62, 63 22 B. ovatus ATCC 8483 64, 65, 66 23 B. caccae ATCC 43185 67, 68, 69 24 C. perfringens ATCC 13124 70, 71, 72 25 C. butyricum ATCC 19398 73, 74, 75 26 C. ramosum ATCC 25582 76, 77, 78 27 C. difficile ATCC 9689 79, 80, 81 28 C. indolis ATCC 25771 82, 83, 84 29 F. russii ATCC 25533 85, 86, 87 30 F. nucleatum ATCC 25586 88, 89, 90 31 B. catenulatum ATCC 27539 91, 92, 93 32 B. angulatum ATCC 27535 94, 95, 96 33 E. rectale ATCC 33656 97, 98, 99 34 E. eligens ATCC 27750 100, 101, 102 35 E. limosum ATCC 8486 103, 104, 105 36 E. lentum ATCC 25553 106, 107, 108 37 L. fermentum ATCC 9338 109, 110, 111 38 E. faecalis ATCC 27274 112, 113, 114 39 P. magnus ATCC 14955 115, 116, 117 40 R. gnavus ATCC 291492 118, 119, 120 Open in a separate window Bacteria and the probe numbers in the microarray 2.2.

Techniques: Bacteria

Journal: Biosensors & bioelectronics

Article Title: Microarray method to monitor 40 intestinal bacterial species in the study of azo dye reduction

doi: 10.1016/j.bios.2004.04.011

Figure Lengend Snippet: Microarray test results read from

Article Snippet: Anaerobic bacteria were cultured at 35 °C in either prereduced anaerobically sterilized (PRAS) Brain Heart Infusion (BIH) broth supplemented with vitamin K and hemin (Remel, Lenexa, KS, USA), inoculated under an oxygen-free cannula using 85% nitrogen, 10% hydrogen and 5% carbon dioxide, or on PRAS brucella blood agar plates supplemented with vitamin K and hemin (Remel). table ft1 table-wrap mode="anchored" t5 caption a7 Number Bacterial species and strain Probe number 1 B. thetaiotaomicron ATCC 29148 1, 2, 3 2 B. vulgatus ATCC 8482 4, 5, 6 3 B. fragilis ATCC 23745 7, 8, 9 4 B. distasonis ATCC 8503 10, 11, 12 5 C. clostridioforme ATCC 29084 13, 14, 15 6 C. leptum ATCC 29065 16, 17, 18 7 F. prausnitzii ATCC 27768 19, 20, 21 8 P. productus ATCC 27340 22, 23, 24 9 R. obeum ATCC 29174 25, 26, 27 10 R. bromii ATCC 27255 28, 29, 30 11 R. callidus ATCC 27760 31, 32, 33 12 R. albus ATCC 27210 34, 35, 36 13 B. longum ATCC 15707 37, 38, 39 14 B. adolescentis ATCC 15703 40, 41, 42 15 B. infantis ATCC 15697 43, 44, 45 16 E. biforme ATCC 27806 46, 47, 48 17 E. aerofaciens ATCC 25986 49, 50, 51 18 L. acidophilus ATCC 4356 52, 53, 54 19 E. coli ATCC 25922 55, 56, 57 20 E. faecium ATCC 19434 58, 59, 60 21 B. uniformis ATCC 8492 61, 62, 63 22 B. ovatus ATCC 8483 64, 65, 66 23 B. caccae ATCC 43185 67, 68, 69 24 C. perfringens ATCC 13124 70, 71, 72 25 C. butyricum ATCC 19398 73, 74, 75 26 C. ramosum ATCC 25582 76, 77, 78 27 C. difficile ATCC 9689 79, 80, 81 28 C. indolis ATCC 25771 82, 83, 84 29 F. russii ATCC 25533 85, 86, 87 30 F. nucleatum ATCC 25586 88, 89, 90 31 B. catenulatum ATCC 27539 91, 92, 93 32 B. angulatum ATCC 27535 94, 95, 96 33 E. rectale ATCC 33656 97, 98, 99 34 E. eligens ATCC 27750 100, 101, 102 35 E. limosum ATCC 8486 103, 104, 105 36 E. lentum ATCC 25553 106, 107, 108 37 L. fermentum ATCC 9338 109, 110, 111 38 E. faecalis ATCC 27274 112, 113, 114 39 P. magnus ATCC 14955 115, 116, 117 40 R. gnavus ATCC 291492 118, 119, 120 Open in a separate window Bacteria and the probe numbers in the microarray 2.2.

Techniques: Microarray

Journal: Biosensors & bioelectronics

Article Title: Microarray method to monitor 40 intestinal bacterial species in the study of azo dye reduction

doi: 10.1016/j.bios.2004.04.011

Figure Lengend Snippet: Azo dye (Direct Blue 15) reduction activity of 17 bacterial species in pure culture

Article Snippet: Anaerobic bacteria were cultured at 35 °C in either prereduced anaerobically sterilized (PRAS) Brain Heart Infusion (BIH) broth supplemented with vitamin K and hemin (Remel, Lenexa, KS, USA), inoculated under an oxygen-free cannula using 85% nitrogen, 10% hydrogen and 5% carbon dioxide, or on PRAS brucella blood agar plates supplemented with vitamin K and hemin (Remel). table ft1 table-wrap mode="anchored" t5 caption a7 Number Bacterial species and strain Probe number 1 B. thetaiotaomicron ATCC 29148 1, 2, 3 2 B. vulgatus ATCC 8482 4, 5, 6 3 B. fragilis ATCC 23745 7, 8, 9 4 B. distasonis ATCC 8503 10, 11, 12 5 C. clostridioforme ATCC 29084 13, 14, 15 6 C. leptum ATCC 29065 16, 17, 18 7 F. prausnitzii ATCC 27768 19, 20, 21 8 P. productus ATCC 27340 22, 23, 24 9 R. obeum ATCC 29174 25, 26, 27 10 R. bromii ATCC 27255 28, 29, 30 11 R. callidus ATCC 27760 31, 32, 33 12 R. albus ATCC 27210 34, 35, 36 13 B. longum ATCC 15707 37, 38, 39 14 B. adolescentis ATCC 15703 40, 41, 42 15 B. infantis ATCC 15697 43, 44, 45 16 E. biforme ATCC 27806 46, 47, 48 17 E. aerofaciens ATCC 25986 49, 50, 51 18 L. acidophilus ATCC 4356 52, 53, 54 19 E. coli ATCC 25922 55, 56, 57 20 E. faecium ATCC 19434 58, 59, 60 21 B. uniformis ATCC 8492 61, 62, 63 22 B. ovatus ATCC 8483 64, 65, 66 23 B. caccae ATCC 43185 67, 68, 69 24 C. perfringens ATCC 13124 70, 71, 72 25 C. butyricum ATCC 19398 73, 74, 75 26 C. ramosum ATCC 25582 76, 77, 78 27 C. difficile ATCC 9689 79, 80, 81 28 C. indolis ATCC 25771 82, 83, 84 29 F. russii ATCC 25533 85, 86, 87 30 F. nucleatum ATCC 25586 88, 89, 90 31 B. catenulatum ATCC 27539 91, 92, 93 32 B. angulatum ATCC 27535 94, 95, 96 33 E. rectale ATCC 33656 97, 98, 99 34 E. eligens ATCC 27750 100, 101, 102 35 E. limosum ATCC 8486 103, 104, 105 36 E. lentum ATCC 25553 106, 107, 108 37 L. fermentum ATCC 9338 109, 110, 111 38 E. faecalis ATCC 27274 112, 113, 114 39 P. magnus ATCC 14955 115, 116, 117 40 R. gnavus ATCC 291492 118, 119, 120 Open in a separate window Bacteria and the probe numbers in the microarray 2.2.

Techniques: Activity Assay

Journal: Science Progress

Article Title: Role of thrombus-derived exosomal lncRNA LOC101928697 in regulating endothelial function via FUS protein interaction in myocardial infarction

doi: 10.1177/00368504251372111

Figure Lengend Snippet: Analysis of the correlation between FUS protein and myocardial infarction. (a) Enrichment Analysis Bar Plot based on differential gene expression profiles in lncRNA microarray analysis.(b) Detection information about lncRNA LOC101928697 binding to FUS proteins in AnnoLnc2 database. (c) Detection information about lncRNA LOC101928697 binding to FUS protein in RBPDP database. (d) Scores in the RPISeq database on the model of lncRNA LOC101928697 binding to FUS protein. (e-g) Prediction information about lncRNA LOC101928697 binding to FUS protein in catRAPID website, (e) Statistical map information about protein and RNA binding sites, (f) Total scoring information, and (g) Interaction map showing the interaction region between protein and RNA. (h-i) Analyses about bioinformatics techniques based on GSE163772 in the GEO database, where (h) is a statistical map of FUS gene expression in endothelial cells of a mouse model of myocardial infarction, and (i) A scatter plot about the correlation between the level of FUS gene expression and the disease state (control vs. myocardial infarction).

Article Snippet: After extensive washing, the bound proteins were eluted, separated by SDS-PAGE, and analyzed by Western blot using anti-FUS antibody (Proteintech, Cat No. 11570-1-AP, dilution 1:5000) to detect the enrichment of FUS protein.

Techniques: Gene Expression, Microarray, Binding Assay, RNA Binding Assay, Control

Journal: Science Progress

Article Title: Role of thrombus-derived exosomal lncRNA LOC101928697 in regulating endothelial function via FUS protein interaction in myocardial infarction

doi: 10.1177/00368504251372111

Figure Lengend Snippet: Interaction of exosomal lncRNA LOC101928697 with FUS proteins. (a and b) The western blot detection of FUS protein expression in each group of cells and the statistical graph. (c) Statistical graph of RT-qPCR to detect the expression of FUS at the mRNA level in each group of cells. (d) The fluorescence graph of fluorescence in situ hybridization (FISH) experiment. In which FUS was labeled with green fluorescence, lncRNA LOC101928697 was labeled with red fluorescence, and the nucleus was labeled with blue fluorescence (20×). (e) Western blot detection of FUS protein following RNA pull-down using sense or antisense LOC101928697 transcripts. (f) Quantification of FUS protein enrichment in sense RNA pull-down versus antisense control, based on densitometric analysis. (g-h) Western blot detection of FUS protein expression in each group of cells after knockdown or overexpression of lncRNA LOC101928697 and the statistical graphs. (i) Statistical graph of mRNA level expression of FUS in each group of cells after knockdown or overexpression of lncRNA LOC101928697 by RT-qPCR assay. a p < 0.05 compared to control group. b p < 0.05 compared to exosome group. c p < 0.05 compared to siRNA + exosome group.

Article Snippet: After extensive washing, the bound proteins were eluted, separated by SDS-PAGE, and analyzed by Western blot using anti-FUS antibody (Proteintech, Cat No. 11570-1-AP, dilution 1:5000) to detect the enrichment of FUS protein.

Techniques: Western Blot, Expressing, Quantitative RT-PCR, Fluorescence, In Situ Hybridization, Labeling, Protein Enrichment, Control, Knockdown, Over Expression

Journal: bioRxiv

Article Title: Senescent cells develop PDK4-dependent hypercatabolism and form an acidic microenvironment to drive cancer resistance

doi: 10.1101/2022.08.29.505761

Figure Lengend Snippet: a. Transcriptome-wide profiling of gene expression changes in primary normal human prostate stromal cell line PSC27 by microarray. Cell lysates were collected for analysis 7 d after treatment. CTRL, control. RAD, radiation. BLEO, bleomycin. HP, hydrogen peroxide. Red highlighted, typical soluble factors of the SASP. Microarray data adapted from Sun et al. with permission from Nature Medicine , copyright 2012, Springer Nature . b. Quantitative RT-PCR to detect PDK4 expression after PSC27 cells were subject to individual treatment as indicated. Cell lysates were collected for measurement 7 d after establishment of stable cell sublines or completion of in vitro treatment. Signals normalized to CTRL. RS, replicative senescence. p16, lentiviral transduction of human tumor suppressor p16 INK4a . RAS, lentiviral transduction of human oncogene HRAS G12V . c. Immunoblot analysis of PDK4 expression in stromal cells as delineated in ( b ). GAPDH, loading control. d. Comparative RT-PCR assay of PDK4 expression after treatment of PSC27 or prostate epithelial cells by agents as indicated. Cell lysates were collected for measurement 7 d after treatment. Signals normalized to CTRL. BPH1, M12, PC3, DU145, LNCaP and VCaP, human epithelial lines of prostate origin. e. Comparative RT-PCR assay of PDK4 expression in human stromal cells 7 d after treatments performed as indicated. WI38, HFL1, HBF1203 and BJ, human stromal lines of different origins. f. A time course RT-PCR assessment of the expression of PDK4 and a set of typical SASP factors (MMP1, WNT16B, SFRP2, SPINK1, MMP3, CXCL8, EREG, ANGPTL4 and AREG) after drug treatment of PSC27 cells in vitro . Numeric numbers indicate the individual days after treatment (indexed at the top line). g. Immunoblot measurement of PDK4 expression at protein level at the individual timepoints as indicated. β-actin, loading control. h. Comparative appraisal of human PDK family expression at transcript level in PSC27 cells after BLEO treatment. Signals normalized to untreated sample per gene. CXCL8, experimental control as a hallmark SASP factor. i. Immunoblot assessment of the expression of PDK4 family members at protein level after BLEO treatment. β-actin, loading control. Data are representative of 3 independent experiments. P values were calculated by Student’s t -test ( b , e , f , h ) and one-way ANOVA ( d ). ^, P > 0.05. *, P < 0.05. **, P < 0.01. ***, P < 0.001. ****, P < 0.0001.

Article Snippet: Human fetal lung stromal lines WI38 and HFL1, and foreskin stromal line BJ were from ATCC and cultured with F-12K medium supplemented with 10% FBS.

Techniques: Gene Expression, Microarray, Control, Quantitative RT-PCR, Expressing, Stable Transfection, In Vitro, Transduction, Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: The American Journal of Pathology

Article Title: Interspecies Comparison of Human and Murine Scleroderma Reveals IL-13 and CCL2 as Disease Subset-Specific Targets

doi: 10.1016/j.ajpath.2011.11.024

Figure Lengend Snippet: The IL-13 pathway is activated in SSc and sclGVHD skin. A: Gene expression data for the 491 IL-13–responsive genes identified in human dermal fibroblasts were extracted from the SSc skin data of Milano et al.4 SSc skin biopsy samples are ordered by intrinsic subset and genes are organized by hierarchical clustering. The centroid average of the IL-13–responsive gene signature at maximal induction (12 and 24 hours) is shown to the left of the heat map. Pearson's correlation coefficients between the centroid and individual patient sample are plotted below each array. B: Expression data for the 734 genes reported by Fulkerson et al26 as IL-13–inducible genes in mouse lung were extracted from the sclGVHD microarray data set obtained 2 weeks and 5 weeks after splenocyte transfer (n = 4 per group). Genes and arrays were organized by hierarchical clustering. The relative expression of IL-13–inducible genes (centroid) in lungs is shown to the left of the heat map. C: Representative IL-13 IHC on skin biopsies obtained from an SSc patient and a normal control subject. Arrowheads indicate IL-13+ cells. Original magnification, ×400. D: Blinded quantification of the number of IHC IL-13+ cells per high-power field (×600) in skin biopsies from SSc patients and normal control subjects. SSc, n = 18; control, n = 6; P = 0.0037. E: IL-13 ELISA on the tissue culture supernatants of skin explants from BALB/c Rag2−/− mice that received either syngeneic BALB/c (n = 3) or allogeneic B10.D2 (n = 3) splenocytes 2 weeks earlier (P = 0.002).

Article Snippet: The 75 microarray human scleroderma data set, previously described, 4 is publicly available at the National Center for Biotechnology Information GEO site ( http://www.ncbi.nlm.nih.gov/geo ; accession {"type":"entrez-geo","attrs":{"text":"GSE9285","term_id":"9285"}} GSE9285 ).

Techniques: Gene Expression, Expressing, Microarray, Control, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Using a Resequencing Microarray as a Multiple Respiratory Pathogen Detection Assay ▿ †

doi: 10.1128/JCM.01870-06

Figure Lengend Snippet: Analytic sensitivity of microarray-based detection for prototype control strains

Article Snippet: Thus, hybridization to a series of probe sets provides redundant presence/absence information for the organism while also revealing strain-specific single-nucleotide polymorphisms relative to the sequence chosen. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Organism a Sample type a Strain Sample source b Detection limit (genome copies) c HAdV-4 d DNA RI-67 ATCC 10 2 HAdV-4 vaccine DNA CL68578 NHRC 10 2 HAdV-4FS_Navy DNA NRHC 10 2 HAdV-4FS_AirForce DNA AFIOH 10 2 HAdV-4FS_AirForce Viral particles ADL 10 2 HAdV-5 DNA Adenoid 75 ATCC 10 3 HAdV-7 DNA Gomen ATCC 10 2 HAdV-7 Viral particles Gomen ATCC ND HAdV-7a vaccine DNA 55142 NHRC 10 2 HAdV-7FS_Navy DNA NHRC 10 3 B. anthracis DNA Ames AFIP 10 1 B. anthracis Bacterial cells Sterne CRP ND B. pertussis DNA NHRC 10 2 Chlamydia pneumoniae DNA ABi 10 1 Influenza A virus (H1N1) Viral particles PR/8/34 ABi 10 2 Influenza A virus (H3N2) RNA ADL 10 2 Influenza A virus (H5N1) RNA AFIOH 10 1 Influenza B virus Viral particles B/Lee/40 ABi 10 3 Francisella tularensis DNA SCHU4 ATCC 10 3 F. tularensis Bacterial cells SCHU4 CRP ND Human coronavirus DNA/RNA 229E ATCC 10 3 Human coronavirus Viral particles 229E ATCC ND Human coronavirus DNA/RNA OC43 ATCC 10 3 Human coronavirus Viral particles OC43 ATCC ND Rhinovirus 89 Viral particles 41467 Gallo ATCC 10 3 Lassa virus e Plasmids BlueHeron 10 3 M. pneumoniae DNA AFIP 10 3 M. pneumoniae Bacterial cells NHRC ND N. meningitidis DNA Murray ATCC 10 2 Parainfluenza virus 1 Viral particles C-35 ATCC 10 3 Parainfluenza virus 3 Viral particles C 243 ATCC 10 3 RSV A Viral particles A-2 ATCC 10 3 RSV B Viral particles B WV/14617/85 ATCC 10 2 S. pneumoniae DNA AFIP 10 2 S. pyogenes DNA Rosenbach ATCC 10 3 S. pyogenes Bacterial cells NHRC ND Variola major virus e Plasmids BlueHeron 10 3 Vaccinia virus DNA Lister ABi 10 3 Y. pestis DNA D27 AFIP 10 3 Ebola virus e Plasmids BlueHeron 10 3 Open in a separate window a Samples were generated by mixing purified nucleic acid templates in Tris-EDTA buffer to create 10 6 genome copies/μl of stock solution.

Techniques: Microarray, Control, Virus

Journal:

Article Title: Using a Resequencing Microarray as a Multiple Respiratory Pathogen Detection Assay ▿ †

doi: 10.1128/JCM.01870-06

Figure Lengend Snippet: Differentiation by RPM v.1 of various HAdVs causing febrile respiratory infections

Article Snippet: Thus, hybridization to a series of probe sets provides redundant presence/absence information for the organism while also revealing strain-specific single-nucleotide polymorphisms relative to the sequence chosen. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Organism a Sample type a Strain Sample source b Detection limit (genome copies) c HAdV-4 d DNA RI-67 ATCC 10 2 HAdV-4 vaccine DNA CL68578 NHRC 10 2 HAdV-4FS_Navy DNA NRHC 10 2 HAdV-4FS_AirForce DNA AFIOH 10 2 HAdV-4FS_AirForce Viral particles ADL 10 2 HAdV-5 DNA Adenoid 75 ATCC 10 3 HAdV-7 DNA Gomen ATCC 10 2 HAdV-7 Viral particles Gomen ATCC ND HAdV-7a vaccine DNA 55142 NHRC 10 2 HAdV-7FS_Navy DNA NHRC 10 3 B. anthracis DNA Ames AFIP 10 1 B. anthracis Bacterial cells Sterne CRP ND B. pertussis DNA NHRC 10 2 Chlamydia pneumoniae DNA ABi 10 1 Influenza A virus (H1N1) Viral particles PR/8/34 ABi 10 2 Influenza A virus (H3N2) RNA ADL 10 2 Influenza A virus (H5N1) RNA AFIOH 10 1 Influenza B virus Viral particles B/Lee/40 ABi 10 3 Francisella tularensis DNA SCHU4 ATCC 10 3 F. tularensis Bacterial cells SCHU4 CRP ND Human coronavirus DNA/RNA 229E ATCC 10 3 Human coronavirus Viral particles 229E ATCC ND Human coronavirus DNA/RNA OC43 ATCC 10 3 Human coronavirus Viral particles OC43 ATCC ND Rhinovirus 89 Viral particles 41467 Gallo ATCC 10 3 Lassa virus e Plasmids BlueHeron 10 3 M. pneumoniae DNA AFIP 10 3 M. pneumoniae Bacterial cells NHRC ND N. meningitidis DNA Murray ATCC 10 2 Parainfluenza virus 1 Viral particles C-35 ATCC 10 3 Parainfluenza virus 3 Viral particles C 243 ATCC 10 3 RSV A Viral particles A-2 ATCC 10 3 RSV B Viral particles B WV/14617/85 ATCC 10 2 S. pneumoniae DNA AFIP 10 2 S. pyogenes DNA Rosenbach ATCC 10 3 S. pyogenes Bacterial cells NHRC ND Variola major virus e Plasmids BlueHeron 10 3 Vaccinia virus DNA Lister ABi 10 3 Y. pestis DNA D27 AFIP 10 3 Ebola virus e Plasmids BlueHeron 10 3 Open in a separate window a Samples were generated by mixing purified nucleic acid templates in Tris-EDTA buffer to create 10 6 genome copies/μl of stock solution.

Techniques:

Journal: Oncotarget

Article Title: Glucose deprivation elicits phenotypic plasticity via ZEB1-mediated expression of NNMT

doi: 10.18632/oncotarget.15429

Figure Lengend Snippet: ( A ) qRT-PCR shows that mRNA expression of ZEB1 is consistently upregulated in all glucose-restricted sublines generated from OVCAR3, OVCAR4 and OAW28 cell lines. ( B ) NNMT expression correlates with the mesenchymal subtype of HGSC patients (M) as defined by the Cancer Genome Atlas and the C1 subtype of HGSC patients, characterized by the reactive stroma gene signature . ( C ) NNMT protein expression is consistently higher in mesenchymal-like ovarian cancer cell lines (low E-cadherin; high vimentin, ZEB1 or SLUG expression: KURAMOCHI, TYKNU, HEY, SKOV3, FUOV1) compared to epithelial-like cell lines (high E-cadherin; low vimentin, ZEB1 or SLUG expression: OVCAR3, OVCA433, OAW28, COV318). However, some epithelial-like cell lines, such as CAOV3, OV90 and OVCAR4, demonstrate relatively high NNMT levels. ( D ) NNMT mRNA expression positively correlates with elevated ZEB1 and Vim (vimentin) expression, and shows a tendency to negatively correlate with CDH1 (E-cadherin) expression in ovarian cancer cell lines. ( E ) Western blot analysis shows that parental OVCAR3 cells ectopically expressing ZEB1 undergo epithelial-to-mesenchymal transition (EMT), as manifested by decreased expression of E-cadherin and increased expression of N-cadherin and vimentin. Overexpression of ZEB1, but not SLUG, induced NNMT protein expression. ( F ) qRT-PCR analysis shows that ZEB1 overexpression in OVCAR3 cells induces NNMT, MMP2, SPARC and decreases CLDN4 and CLDN7 expression, but has no effect on the SLC2A1 and G6PD expression. ( G ) Biolog Microarray studies shows that ectopic ZEB1 expression recapitulates metabolic adaptations observed in OVCAR3 glucose-restricted cells, such as increased utilization of sugars (D-galactose, dextrin, maltotriose, xylitol, D-fructose), ketones (D, L-β-hydroxy-butyric acid), D, L-lactic acid and methylated substrates (α-methyl-D-galactoside, α-methyl-D-glucoside, mono-methyl succinate) in the absence of glucose. Asterisks (*) denote substrates differentially utilized between control and transformed cells demonstrating statistical significance P < 0.05. For all figure panels, statistical calculations were performed using a two-tailed Student's t -test (* 0.001 < P < 0.05; ** P < 0.001).

Article Snippet: Human ovarian carcinoma cell lines CAOV3, OVCAR3, OV90 and SKOV3 were obtained from ATCC (Manassas, VA).

Techniques: Quantitative RT-PCR, Expressing, Generated, Western Blot, Over Expression, Microarray, Methylation, Control, Transformation Assay, Two Tailed Test