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  • 97
    Thermo Fisher gene exp maf mm02581355 s1
    PLZF regulates the expression of <t>c-Maf</t> . Expression of Maf was analyzed by TaqMan qPCR in sorted total thymic, splenic, and liver iNKT (TCRβ + CD1d-Tet + ) and γδNKT (TCRγδ + Vδ6.3 + ) cells (A) or in stage 0/stage 1 iNKT cells (CD44 − NK1.1 − TCRβ + CD1d-Tet + ) (B) from mixed bm chimeras. Mean relative c-Maf expression normalized against GAPDH expression is shown. Error bars represent SD (individual mice). Statistical significance is indicated where reached. iNKT cell were sorted from thymi (C) and spleens (D,E) of WT or PLZF lu/lu mice, cultured for 2 days in the presence of IL-7 and IL-15, infected with IRES-GFP or c-Maf-IRES-GFP retroviruses and stimulated with PMA/Ionomycin 2 days after infection. Expression of IFNγ and IL-4 in GFP + cells (C,D) as well as IL-10 and GFP in all NKT cells (E) is shown. Representative FACS plots from one of two (C) and one of three (D,E) experiments are shown. Histogram overlays comparing cytokine production by wt and lu/lu cells in response to c-Maf expression are shown for (D) .
    Gene Exp Maf Mm02581355 S1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 67 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Illumina Inc minor allele frequency maf
    PLZF regulates the expression of <t>c-Maf</t> . Expression of Maf was analyzed by TaqMan qPCR in sorted total thymic, splenic, and liver iNKT (TCRβ + CD1d-Tet + ) and γδNKT (TCRγδ + Vδ6.3 + ) cells (A) or in stage 0/stage 1 iNKT cells (CD44 − NK1.1 − TCRβ + CD1d-Tet + ) (B) from mixed bm chimeras. Mean relative c-Maf expression normalized against GAPDH expression is shown. Error bars represent SD (individual mice). Statistical significance is indicated where reached. iNKT cell were sorted from thymi (C) and spleens (D,E) of WT or PLZF lu/lu mice, cultured for 2 days in the presence of IL-7 and IL-15, infected with IRES-GFP or c-Maf-IRES-GFP retroviruses and stimulated with PMA/Ionomycin 2 days after infection. Expression of IFNγ and IL-4 in GFP + cells (C,D) as well as IL-10 and GFP in all NKT cells (E) is shown. Representative FACS plots from one of two (C) and one of three (D,E) experiments are shown. Histogram overlays comparing cytokine production by wt and lu/lu cells in response to c-Maf expression are shown for (D) .
    Minor Allele Frequency Maf, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 91/100, based on 170 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Santa Cruz Biotechnology c maf
    IRF4 assembles into a complex with <t>AhR</t> in the ICOS promoter in human act-A–iTr1 cells. Schematic representation of the human IL10 promoter ( A ) and ICOS locus ( B ); the <t>c-Maf,</t> AhR, and IRF4 putative binding sites are shown. Positions are given in
    C Maf, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 235 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    c maf  (4Gene)
    91
    4Gene c maf
    <t>c-Maf</t> and IL-10 induction by <t>Th17-polarizing</t> signals are independent of IL-4/Stat6/GATA-3. A , Intracellular staining for IL-4, IL-10, and ELISA for IL-10 in wild-type, Stat6-deficient, T-bet-deficient, or Stat6/T-bet double-deficient CD4 + CD25 –
    C Maf, supplied by 4Gene, used in various techniques. Bioz Stars score: 91/100, based on 216 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    4Gene linc maf 4
    <t>Linc-MAF-4</t> contributes to T H 1 cell differentiation ( a ) Expression of linc-MAF-4 and MAF assessed at different time points by RT-qPCR in activated CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions (average of four technical replicates ± SEM). See also Supplementary Fig. 4 b,c . ( b ) ChIP-qPCR analysis of H3K4me3 and RNA polymerase II occupancy at MAF locus in CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions at day 8 post-activation. Enrichment is a percentage of input (average of at least 5 independent experiments ± SEM). One-tailed t -test * P
    Linc Maf 4, supplied by 4Gene, used in various techniques. Bioz Stars score: 88/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Illumina Inc maf
    Power for the W2WK (red), traditional joint (blue), and traditional main effect (green) approaches by causal <t>SNP</t> under the M1 model. An “x” marks the 14 SNPs that were modeled as genotyped in our simulations. The <t>MAF</t> of each SNP (grey line) is along the right Y-axis. Plot on the left assumes an additive model; the plot on the right assumes the underlying model is dominant, but was tested as additive. Inset plot shows the underlying model (M1): solid black line represents genotypic effect among individuals who have been exposed to the environmental insult. Dotted black line shows genotypic effect of individuals who were not exposed to the environmental insult under the M1 model. Dotted grey lines indicate alternate models that are considered elsewhere in this manuscript.
    Maf, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 92/100, based on 136 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Santa Cruz Biotechnology anti c maf antibody
    Sox5 and <t>c-Maf</t> activate the promoter of RORγt gene. (A, top) VISTA plot of CNSs of Rorc gene locus. (middle panels) Naive CD4 + T cells from CD4 cre Sox5 fl/fl or Sox5 fl/fl mice were stimulated under Th17 conditions for 8 h. ChIP-qPCR assay for CNS of Rorc gene locus was performed with <t>anti–c-Maf</t> antibody, anti-p300 antibody, or control rabbit IgG. (bottom) Naive CD4 + T cells from WT mice were stimulated under Th17 conditions for 8 h. ChIP-qPCR assay for CNS of Rorc gene locus was performed with anti-Stat3 antibody, anti-H3K4me3 antibody, or control rabbit IgG. Results are expressed as the percent input for each ChIP fraction. Data are representative of three independent experiments. (B) Luciferase assay of indicated RORγt-Luci in EL4 cells. Data are compiled from three independent experiments. (C) Sequence of RORγt promoter (from −1648 to −1448). Putative Sox5 binding site is indicated by red and putative MAREs are indicated by blue. Mutated sequences are shown in lower lines. (D, top) Representative data of DNA precipitation assay of HA-Sox5 with biotinylated double-stranded DNA probes containing a portion of RORγt promoter or its mutant at Sox5-binding site. As a positive control (PC), FXO+ probe, which harbors two Sox consensus sites, was used. (bottom) Representative data of DNA-precipitation assay of c-Maf with biotinylated double-stranded DNA probes containing a portion of RORγt promoter or its mutants at MARE sites. Shown are data representative of three independent experiments. (E) Naive CD4 + T cells were co-infected with retroviruses of either pMX-IN or pMX-IN-Sox5t and with those of either MIT or MIT-c-Maf for 24 h. Cells were reinfected with WT-RORγtP for 24 h, and GFP versus hCD4 in NGFR + Thy1.1 + cells was evaluated by flow cytometry. (F) Naive CD4 + T cells were co-infected with retroviruses of indicated vectors and analyzed as described in Fig. 7 E . (G) Naive CD4 + T cells from Sox5 fl/fl mice and CD4 cre Sox5 fl/fl mice were stimulated in neutral conditions for 48 h and infected with retroviruses of hCD4-pA-GFP (empty), WT-RORγtP, or ΔSox5-ΔMARE-RORγtP under Th17 conditions for additional 24 h. The expression levels of GFP were evaluated as described in Fig. 7 E . Data shown are representative of three independent experiments.
    Anti C Maf Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 88/100, based on 125 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher c maf
    Highly efficient target gene KO in nonactivated human and mouse primary T cells with an optimized CRISPR/Cas9 RNP transfection approach. (A–D) KO efficiency by flow cytometry of CXCR4, CD127, and CCR7 in nonactivated human CD4 T cells 72 h after transfection (A); PD1, TIGIT, and CTLA4 in nonactivated human CD8 T cells 72 h after transfection followed by 72 h of stimulation with anti-CD3/anti-CD28 (B); CD90 and CTLA4 in IL-7–preconditioned nonactivated mouse CD4 T cells 5 d after transfection and incubation with IL-7 (C); and CD8α and CTLA4 in IL-7–preconditioned nonactivated mouse CD8 + T cells after transfection and 5 d of incubation in the presence of IL-7 followed by 48 h stimulation with anti-CD3/anti-CD28 (D; all compared with nontargeting control Cas9 RNP-transfected T cells). Data are presented as mean ± SD ( n = 2) and representative of three (A) or two (B–D) independent experiments. (E) IFN-γ expression by flow cytometry in crIFNγ or NTC-transfected nonactivated human CD8 + T cells cultured for 3 d and restimulated for 4 h with PMA/ionomycin. Data are presented as mean ± SD ( n = 2) and representative of two independent experiments. (F) <t>FoxP3</t> and <t>c-Maf</t> expression by flow cytometry in crFoxp3 or NTC transfected IL-7–preconditioned nonactivated mouse CD4 + T cells 5 d after transfection with IL-7 followed by 3 d of polarization into iTreg with TGF-β and IL-2. Data are presented as mean ± SD ( n = 2) and representative of two independent experiments. (G–I) KO efficiency by flow cytometry of double KOs targeting CD127 and CCR7 in nonactivated human CD4 + T cells 72 h after transfection (G), CD90 and CTLA4 in nonactivated mouse CD8 T cells after transfection and 5 d of incubation in the presence of IL-7 followed by 48 h stimulation with anti-CD3/anti-CD28 (H), or CD90 and FoxP3 in nonactivated mouse CD4 + T cells after transfection and 5 d of incubation with IL-7 followed by 3 d of polarization into iTreg with TGF-β and IL-2 with anti-CD3/anti-CD28 (I). Data are presented as mean ± SD ( n = 2) and representative of two independent experiments. ***, P
    C Maf, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Bethyl rabbit anti c maf
    Expression of <t>c-Maf</t> and its target genes in the dorsal horn of MafA −/− mice. A , B , Immunostaining of c-Maf was performed on sections of spinal cord in MafA +/− and MafA −/− mice at P0. Insets show the <t>anti-c-Maf</t> immunofluorescence signal in the DRG of control and MafA deletion mice. Note comparable expression of c-Maf in both the dorsal horn and the DRG in MafA −/− versus control mice. C–H , In situ hybridization was performed with Gabra5 ( C , D ), CCK ( E , F ), and Rora ( G , H ) probes on sections of cervical/thoracic spinal cord in MafA +/− and MafA −/− mice at P0. No apparent difference was observed in MafA mutant and control mice. Scale bars: (in B , H ) A–H , 50 μm; (in B , inset), A , B , insets, 50 μm.
    Rabbit Anti C Maf, supplied by Bethyl, used in various techniques. Bioz Stars score: 92/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Pfizer Inc maf
    Expression of <t>c-Maf</t> and its target genes in the dorsal horn of MafA −/− mice. A , B , Immunostaining of c-Maf was performed on sections of spinal cord in MafA +/− and MafA −/− mice at P0. Insets show the <t>anti-c-Maf</t> immunofluorescence signal in the DRG of control and MafA deletion mice. Note comparable expression of c-Maf in both the dorsal horn and the DRG in MafA −/− versus control mice. C–H , In situ hybridization was performed with Gabra5 ( C , D ), CCK ( E , F ), and Rora ( G , H ) probes on sections of cervical/thoracic spinal cord in MafA +/− and MafA −/− mice at P0. No apparent difference was observed in MafA mutant and control mice. Scale bars: (in B , H ) A–H , 50 μm; (in B , inset), A , B , insets, 50 μm.
    Maf, supplied by Pfizer Inc, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Thermo Fisher gene exp maf hs00193519 m1
    <t>Linc-MAF-4</t> contributes to T H 1 cell differentiation ( a ) Expression of linc-MAF-4 and MAF assessed at different time points by RT-qPCR in activated CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions (average of four technical replicates ± SEM). See also Supplementary Fig. 4 b,c . ( b ) ChIP-qPCR analysis of H3K4me3 and RNA polymerase II occupancy at MAF locus in CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions at day 8 post-activation. Enrichment is a percentage of input (average of at least 5 independent experiments ± SEM). One-tailed t -test * P
    Gene Exp Maf Hs00193519 M1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam anti maf antibody
    <t>Linc-MAF-4</t> contributes to T H 1 cell differentiation ( a ) Expression of linc-MAF-4 and MAF assessed at different time points by RT-qPCR in activated CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions (average of four technical replicates ± SEM). See also Supplementary Fig. 4 b,c . ( b ) ChIP-qPCR analysis of H3K4me3 and RNA polymerase II occupancy at MAF locus in CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions at day 8 post-activation. Enrichment is a percentage of input (average of at least 5 independent experiments ± SEM). One-tailed t -test * P
    Anti Maf Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Schiff Nutrition International hla maf
    <t>Linc-MAF-4</t> contributes to T H 1 cell differentiation ( a ) Expression of linc-MAF-4 and MAF assessed at different time points by RT-qPCR in activated CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions (average of four technical replicates ± SEM). See also Supplementary Fig. 4 b,c . ( b ) ChIP-qPCR analysis of H3K4me3 and RNA polymerase II occupancy at MAF locus in CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions at day 8 post-activation. Enrichment is a percentage of input (average of at least 5 independent experiments ± SEM). One-tailed t -test * P
    Hla Maf, supplied by Schiff Nutrition International, used in various techniques. Bioz Stars score: 88/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher gene exp maf hs04185012 s1
    Cholesterol biosynthesis pathway inhibition interferes with <t>c-Maf</t> expression. Purified human CD4 + T cells stimulated in vitro with plate-bound α-CD3 (2 μgml −1 ) + α-CD46 (5 μgml −1 ) and recombinant human interleukin-2 (rhIL-2) (50 Uml −1 ) were cultured for 36 h in the presence of 25-hydroxycholesterol (25-HC). a Expression levels of MAF mRNA ( n = 7). b Expression levels of PRDM1 mRNA ( n = 6). c Representative flow cytometric analysis of intracellular c-Maf staining. d Normalised frequency of c-Maf + cells ( n = 16). e Normalised frequency of c-Maf + cells cultured under fully supplemented medium (M) and cholesterol (500×) (Ch) in the presence or absence of 2μM 25-HC ( n = 3). f Correlation between frequency of c-Maf + and IL-10 + cells, numbers denote r and p values for Spearman's correlation test. Graphs show independent donors (dots) normalised to untreated cells; bars represent median values. *
    Gene Exp Maf Hs04185012 S1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    4Gene transcription factor c maf
    Cholesterol biosynthesis pathway inhibition interferes with <t>c-Maf</t> expression. Purified human CD4 + T cells stimulated in vitro with plate-bound α-CD3 (2 μgml −1 ) + α-CD46 (5 μgml −1 ) and recombinant human interleukin-2 (rhIL-2) (50 Uml −1 ) were cultured for 36 h in the presence of 25-hydroxycholesterol (25-HC). a Expression levels of MAF mRNA ( n = 7). b Expression levels of PRDM1 mRNA ( n = 6). c Representative flow cytometric analysis of intracellular c-Maf staining. d Normalised frequency of c-Maf + cells ( n = 16). e Normalised frequency of c-Maf + cells cultured under fully supplemented medium (M) and cholesterol (500×) (Ch) in the presence or absence of 2μM 25-HC ( n = 3). f Correlation between frequency of c-Maf + and IL-10 + cells, numbers denote r and p values for Spearman's correlation test. Graphs show independent donors (dots) normalised to untreated cells; bars represent median values. *
    Transcription Factor C Maf, supplied by 4Gene, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Assurex Health maf
    Cholesterol biosynthesis pathway inhibition interferes with <t>c-Maf</t> expression. Purified human CD4 + T cells stimulated in vitro with plate-bound α-CD3 (2 μgml −1 ) + α-CD46 (5 μgml −1 ) and recombinant human interleukin-2 (rhIL-2) (50 Uml −1 ) were cultured for 36 h in the presence of 25-hydroxycholesterol (25-HC). a Expression levels of MAF mRNA ( n = 7). b Expression levels of PRDM1 mRNA ( n = 6). c Representative flow cytometric analysis of intracellular c-Maf staining. d Normalised frequency of c-Maf + cells ( n = 16). e Normalised frequency of c-Maf + cells cultured under fully supplemented medium (M) and cholesterol (500×) (Ch) in the presence or absence of 2μM 25-HC ( n = 3). f Correlation between frequency of c-Maf + and IL-10 + cells, numbers denote r and p values for Spearman's correlation test. Graphs show independent donors (dots) normalised to untreated cells; bars represent median values. *
    Maf, supplied by Assurex Health, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Data MATRIX maf
    Cholesterol biosynthesis pathway inhibition interferes with <t>c-Maf</t> expression. Purified human CD4 + T cells stimulated in vitro with plate-bound α-CD3 (2 μgml −1 ) + α-CD46 (5 μgml −1 ) and recombinant human interleukin-2 (rhIL-2) (50 Uml −1 ) were cultured for 36 h in the presence of 25-hydroxycholesterol (25-HC). a Expression levels of MAF mRNA ( n = 7). b Expression levels of PRDM1 mRNA ( n = 6). c Representative flow cytometric analysis of intracellular c-Maf staining. d Normalised frequency of c-Maf + cells ( n = 16). e Normalised frequency of c-Maf + cells cultured under fully supplemented medium (M) and cholesterol (500×) (Ch) in the presence or absence of 2μM 25-HC ( n = 3). f Correlation between frequency of c-Maf + and IL-10 + cells, numbers denote r and p values for Spearman's correlation test. Graphs show independent donors (dots) normalised to untreated cells; bars represent median values. *
    Maf, supplied by Data MATRIX, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher maf
    Cholesterol biosynthesis pathway inhibition interferes with <t>c-Maf</t> expression. Purified human CD4 + T cells stimulated in vitro with plate-bound α-CD3 (2 μgml −1 ) + α-CD46 (5 μgml −1 ) and recombinant human interleukin-2 (rhIL-2) (50 Uml −1 ) were cultured for 36 h in the presence of 25-hydroxycholesterol (25-HC). a Expression levels of MAF mRNA ( n = 7). b Expression levels of PRDM1 mRNA ( n = 6). c Representative flow cytometric analysis of intracellular c-Maf staining. d Normalised frequency of c-Maf + cells ( n = 16). e Normalised frequency of c-Maf + cells cultured under fully supplemented medium (M) and cholesterol (500×) (Ch) in the presence or absence of 2μM 25-HC ( n = 3). f Correlation between frequency of c-Maf + and IL-10 + cells, numbers denote r and p values for Spearman's correlation test. Graphs show independent donors (dots) normalised to untreated cells; bars represent median values. *
    Maf, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 47 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam avian musculoaponeurotic fibrosarcoma virus oncogene c maf antibody
    Cholesterol biosynthesis pathway inhibition interferes with <t>c-Maf</t> expression. Purified human CD4 + T cells stimulated in vitro with plate-bound α-CD3 (2 μgml −1 ) + α-CD46 (5 μgml −1 ) and recombinant human interleukin-2 (rhIL-2) (50 Uml −1 ) were cultured for 36 h in the presence of 25-hydroxycholesterol (25-HC). a Expression levels of MAF mRNA ( n = 7). b Expression levels of PRDM1 mRNA ( n = 6). c Representative flow cytometric analysis of intracellular c-Maf staining. d Normalised frequency of c-Maf + cells ( n = 16). e Normalised frequency of c-Maf + cells cultured under fully supplemented medium (M) and cholesterol (500×) (Ch) in the presence or absence of 2μM 25-HC ( n = 3). f Correlation between frequency of c-Maf + and IL-10 + cells, numbers denote r and p values for Spearman's correlation test. Graphs show independent donors (dots) normalised to untreated cells; bars represent median values. *
    Avian Musculoaponeurotic Fibrosarcoma Virus Oncogene C Maf Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher anti c maf efluor660
    Cholesterol biosynthesis pathway inhibition interferes with <t>c-Maf</t> expression. Purified human CD4 + T cells stimulated in vitro with plate-bound α-CD3 (2 μgml −1 ) + α-CD46 (5 μgml −1 ) and recombinant human interleukin-2 (rhIL-2) (50 Uml −1 ) were cultured for 36 h in the presence of 25-hydroxycholesterol (25-HC). a Expression levels of MAF mRNA ( n = 7). b Expression levels of PRDM1 mRNA ( n = 6). c Representative flow cytometric analysis of intracellular c-Maf staining. d Normalised frequency of c-Maf + cells ( n = 16). e Normalised frequency of c-Maf + cells cultured under fully supplemented medium (M) and cholesterol (500×) (Ch) in the presence or absence of 2μM 25-HC ( n = 3). f Correlation between frequency of c-Maf + and IL-10 + cells, numbers denote r and p values for Spearman's correlation test. Graphs show independent donors (dots) normalised to untreated cells; bars represent median values. *
    Anti C Maf Efluor660, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PLZF regulates the expression of c-Maf . Expression of Maf was analyzed by TaqMan qPCR in sorted total thymic, splenic, and liver iNKT (TCRβ + CD1d-Tet + ) and γδNKT (TCRγδ + Vδ6.3 + ) cells (A) or in stage 0/stage 1 iNKT cells (CD44 − NK1.1 − TCRβ + CD1d-Tet + ) (B) from mixed bm chimeras. Mean relative c-Maf expression normalized against GAPDH expression is shown. Error bars represent SD (individual mice). Statistical significance is indicated where reached. iNKT cell were sorted from thymi (C) and spleens (D,E) of WT or PLZF lu/lu mice, cultured for 2 days in the presence of IL-7 and IL-15, infected with IRES-GFP or c-Maf-IRES-GFP retroviruses and stimulated with PMA/Ionomycin 2 days after infection. Expression of IFNγ and IL-4 in GFP + cells (C,D) as well as IL-10 and GFP in all NKT cells (E) is shown. Representative FACS plots from one of two (C) and one of three (D,E) experiments are shown. Histogram overlays comparing cytokine production by wt and lu/lu cells in response to c-Maf expression are shown for (D) .

    Journal: Frontiers in Immunology

    Article Title: PLZF Controls the Expression of a Limited Number of Genes Essential for NKT Cell Function

    doi: 10.3389/fimmu.2012.00374

    Figure Lengend Snippet: PLZF regulates the expression of c-Maf . Expression of Maf was analyzed by TaqMan qPCR in sorted total thymic, splenic, and liver iNKT (TCRβ + CD1d-Tet + ) and γδNKT (TCRγδ + Vδ6.3 + ) cells (A) or in stage 0/stage 1 iNKT cells (CD44 − NK1.1 − TCRβ + CD1d-Tet + ) (B) from mixed bm chimeras. Mean relative c-Maf expression normalized against GAPDH expression is shown. Error bars represent SD (individual mice). Statistical significance is indicated where reached. iNKT cell were sorted from thymi (C) and spleens (D,E) of WT or PLZF lu/lu mice, cultured for 2 days in the presence of IL-7 and IL-15, infected with IRES-GFP or c-Maf-IRES-GFP retroviruses and stimulated with PMA/Ionomycin 2 days after infection. Expression of IFNγ and IL-4 in GFP + cells (C,D) as well as IL-10 and GFP in all NKT cells (E) is shown. Representative FACS plots from one of two (C) and one of three (D,E) experiments are shown. Histogram overlays comparing cytokine production by wt and lu/lu cells in response to c-Maf expression are shown for (D) .

    Article Snippet: Mm02581355_s1 (Maf), Mm00711781_m1 (Id2), and Mm99999915_g1 (GAPDH) TaqMan Gene Expression Assays (Applied Biosystems) were used.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Mouse Assay, Cell Culture, Infection, FACS

    IRF4 assembles into a complex with AhR in the ICOS promoter in human act-A–iTr1 cells. Schematic representation of the human IL10 promoter ( A ) and ICOS locus ( B ); the c-Maf, AhR, and IRF4 putative binding sites are shown. Positions are given in

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Activin-A co-opts IRF4 and AhR signaling to induce human regulatory T cells that restrain asthmatic responses

    doi: 10.1073/pnas.1616942114

    Figure Lengend Snippet: IRF4 assembles into a complex with AhR in the ICOS promoter in human act-A–iTr1 cells. Schematic representation of the human IL10 promoter ( A ) and ICOS locus ( B ); the c-Maf, AhR, and IRF4 putative binding sites are shown. Positions are given in

    Article Snippet: Slides were incubated overnight at 4 °C with primary biotinylated antibodies against human AhR (Santa Cruz Biotechnology), c-Maf (Santa Cruz Biotechnology), IRF4 (R & D Systems), or control IgG (R & D Systems).

    Techniques: Activated Clotting Time Assay, Binding Assay

    IRF4, AhR, and c-Maf are activated upon stimulation of human CD4 + T cells with activin-A. ( A ) Human naive CD4 + T cells were stimulated with allergen-loaded, mitomycin-treated APCs in the presence of PBS or activin-A for 3 d. Real-time PCR analysis of

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Activin-A co-opts IRF4 and AhR signaling to induce human regulatory T cells that restrain asthmatic responses

    doi: 10.1073/pnas.1616942114

    Figure Lengend Snippet: IRF4, AhR, and c-Maf are activated upon stimulation of human CD4 + T cells with activin-A. ( A ) Human naive CD4 + T cells were stimulated with allergen-loaded, mitomycin-treated APCs in the presence of PBS or activin-A for 3 d. Real-time PCR analysis of

    Article Snippet: Slides were incubated overnight at 4 °C with primary biotinylated antibodies against human AhR (Santa Cruz Biotechnology), c-Maf (Santa Cruz Biotechnology), IRF4 (R & D Systems), or control IgG (R & D Systems).

    Techniques: Real-time Polymerase Chain Reaction

    Kinetin riboside causes suppression of cyclins D1 and D2 and caspase activation in human MM. ( A ) Immunoblot showing suppression of cyclins D1 and D2, but not D3, and induction of caspase-9 cleavage by kinetin riboside (10 μM) at 16 hours in HMCL. Kinetin riboside effects are compared with those of an unrelated control cytotoxic β-lapachone (1 μM, approximately 4 × IC 50 ) or with DMSO vehicle. For visualization, the JJN3 cyclin D2 blot was exposed approximately 2-fold longer than cyclin D2 blots for H929, KMS11, and U266. ( B ) Kinetin riboside (10 μM) induces similar cyclin D1 and D2 suppression and caspase cleavage in CD138-purified primary myeloma cells at 16 hours, shown by immunoblotting. A plasma cell leukemia sample (patient E) was unaffected. The cytogenetics status of the tumor cells or IGH gene translocation is shown, if known. ( C ) Cyclin D and MAF protein levels in H929 cells after exposure to kinetin riboside (10 μM), showing suppression of cyclin D1 and D2 within 6 hours.

    Journal: The Journal of Clinical Investigation

    Article Title: Identification of kinetin riboside as a repressor of CCND1 and CCND2 with preclinical antimyeloma activity

    doi: 10.1172/JCI34149

    Figure Lengend Snippet: Kinetin riboside causes suppression of cyclins D1 and D2 and caspase activation in human MM. ( A ) Immunoblot showing suppression of cyclins D1 and D2, but not D3, and induction of caspase-9 cleavage by kinetin riboside (10 μM) at 16 hours in HMCL. Kinetin riboside effects are compared with those of an unrelated control cytotoxic β-lapachone (1 μM, approximately 4 × IC 50 ) or with DMSO vehicle. For visualization, the JJN3 cyclin D2 blot was exposed approximately 2-fold longer than cyclin D2 blots for H929, KMS11, and U266. ( B ) Kinetin riboside (10 μM) induces similar cyclin D1 and D2 suppression and caspase cleavage in CD138-purified primary myeloma cells at 16 hours, shown by immunoblotting. A plasma cell leukemia sample (patient E) was unaffected. The cytogenetics status of the tumor cells or IGH gene translocation is shown, if known. ( C ) Cyclin D and MAF protein levels in H929 cells after exposure to kinetin riboside (10 μM), showing suppression of cyclin D1 and D2 within 6 hours.

    Article Snippet: PVDF membranes were probed with antibodies against cyclin D1 (DCS-6; Biosource), cyclin D2 (2924; Cell Signaling Technology), cyclin D3 (DCS-22; BioSource), MAF (M-153; Santa Cruz Biotechnology Inc.), cleaved caspase-9 (Asp315, 9505; Cell Signaling Technology), β-actin (4967; Cell Signaling Technology), and α-tubulin (B-7; Santa Cruz Biotechnology Inc.).

    Techniques: Activation Assay, Purification, Translocation Assay

    c-Maf and IL-10 induction by Th17-polarizing signals are independent of IL-4/Stat6/GATA-3. A , Intracellular staining for IL-4, IL-10, and ELISA for IL-10 in wild-type, Stat6-deficient, T-bet-deficient, or Stat6/T-bet double-deficient CD4 + CD25 –

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: c-Maf Regulates IL-10 Expression during Th17 Polarization 1

    doi: 10.4049/jimmunol.0900123

    Figure Lengend Snippet: c-Maf and IL-10 induction by Th17-polarizing signals are independent of IL-4/Stat6/GATA-3. A , Intracellular staining for IL-4, IL-10, and ELISA for IL-10 in wild-type, Stat6-deficient, T-bet-deficient, or Stat6/T-bet double-deficient CD4 + CD25 –

    Article Snippet: Real-time RT-PCR for c-Maf showed that IL-6 combined with TGF β induced similarly high levels of c-Maf in wild-type, Stat6, T-bet, or Stat6/T-bet double-deficient CD4 T cells , indicating that c-Maf expression during Th17 polarization was also independent of Th1 or Th2 signals. c-Maf activates IL-4 gene transcription ( ); however, there was very little IL-4 expression during Th17 polarization despite high levels of c-Maf ( ).

    Techniques: Staining, Enzyme-linked Immunosorbent Assay

    c-Maf and IL-10 induction during Th17 polarization is independent of Stat1. A , Real-time RT-PCR for IL-10, c-Maf, and GATA-3 expression in wild-type and Stat1-deficient CD4 + CD25 – T cells 48 h after anti-CD3 (1 μ g/ml), IL-2 (1 ng/ml), T-depleted

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: c-Maf Regulates IL-10 Expression during Th17 Polarization 1

    doi: 10.4049/jimmunol.0900123

    Figure Lengend Snippet: c-Maf and IL-10 induction during Th17 polarization is independent of Stat1. A , Real-time RT-PCR for IL-10, c-Maf, and GATA-3 expression in wild-type and Stat1-deficient CD4 + CD25 – T cells 48 h after anti-CD3 (1 μ g/ml), IL-2 (1 ng/ml), T-depleted

    Article Snippet: Real-time RT-PCR for c-Maf showed that IL-6 combined with TGF β induced similarly high levels of c-Maf in wild-type, Stat6, T-bet, or Stat6/T-bet double-deficient CD4 T cells , indicating that c-Maf expression during Th17 polarization was also independent of Th1 or Th2 signals. c-Maf activates IL-4 gene transcription ( ); however, there was very little IL-4 expression during Th17 polarization despite high levels of c-Maf ( ).

    Techniques: Quantitative RT-PCR, Expressing

    c-Maf expression correlates with IL-10 production. A , Wild-type CD4 + CD25 – T cells were stimulated with anti-CD3 (1 μ g/ml), IL-2 (1 ng/ml), T-depleted wild-type splenocytes, and IL-6 (10 ng/ml) with variable concentrations of TGF β

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: c-Maf Regulates IL-10 Expression during Th17 Polarization 1

    doi: 10.4049/jimmunol.0900123

    Figure Lengend Snippet: c-Maf expression correlates with IL-10 production. A , Wild-type CD4 + CD25 – T cells were stimulated with anti-CD3 (1 μ g/ml), IL-2 (1 ng/ml), T-depleted wild-type splenocytes, and IL-6 (10 ng/ml) with variable concentrations of TGF β

    Article Snippet: Real-time RT-PCR for c-Maf showed that IL-6 combined with TGF β induced similarly high levels of c-Maf in wild-type, Stat6, T-bet, or Stat6/T-bet double-deficient CD4 T cells , indicating that c-Maf expression during Th17 polarization was also independent of Th1 or Th2 signals. c-Maf activates IL-4 gene transcription ( ); however, there was very little IL-4 expression during Th17 polarization despite high levels of c-Maf ( ).

    Techniques: Expressing

    IL-6 plus TGF β induce high levels of c-Maf expression. A , Real-time RT-PCR for c-Maf, IL-10, ROR γ T, and IL-17 expression in Stat6/T-bet DKO and wild-type CD4 + CD25 – T cells stimulated with anti-CD3 (1 μ g/ml), IL-2 (1 ng/ml),

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: c-Maf Regulates IL-10 Expression during Th17 Polarization 1

    doi: 10.4049/jimmunol.0900123

    Figure Lengend Snippet: IL-6 plus TGF β induce high levels of c-Maf expression. A , Real-time RT-PCR for c-Maf, IL-10, ROR γ T, and IL-17 expression in Stat6/T-bet DKO and wild-type CD4 + CD25 – T cells stimulated with anti-CD3 (1 μ g/ml), IL-2 (1 ng/ml),

    Article Snippet: Real-time RT-PCR for c-Maf showed that IL-6 combined with TGF β induced similarly high levels of c-Maf in wild-type, Stat6, T-bet, or Stat6/T-bet double-deficient CD4 T cells , indicating that c-Maf expression during Th17 polarization was also independent of Th1 or Th2 signals. c-Maf activates IL-4 gene transcription ( ); however, there was very little IL-4 expression during Th17 polarization despite high levels of c-Maf ( ).

    Techniques: Expressing, Quantitative RT-PCR

    c-Maf induces IL-10 production in T cells. A , Intracellular staining for IL-10 and IL-4 expression in wild-type and Stat6-deficient CD4 + CD25 – or CD8 T cells infected with GFP or c-Maf-GFP retrovirus. GFP + cells were isolated after 2 days of stimulation,

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: c-Maf Regulates IL-10 Expression during Th17 Polarization 1

    doi: 10.4049/jimmunol.0900123

    Figure Lengend Snippet: c-Maf induces IL-10 production in T cells. A , Intracellular staining for IL-10 and IL-4 expression in wild-type and Stat6-deficient CD4 + CD25 – or CD8 T cells infected with GFP or c-Maf-GFP retrovirus. GFP + cells were isolated after 2 days of stimulation,

    Article Snippet: Real-time RT-PCR for c-Maf showed that IL-6 combined with TGF β induced similarly high levels of c-Maf in wild-type, Stat6, T-bet, or Stat6/T-bet double-deficient CD4 T cells , indicating that c-Maf expression during Th17 polarization was also independent of Th1 or Th2 signals. c-Maf activates IL-4 gene transcription ( ); however, there was very little IL-4 expression during Th17 polarization despite high levels of c-Maf ( ).

    Techniques: Staining, Expressing, Infection, Isolation

    Linc-MAF-4 contributes to T H 1 cell differentiation ( a ) Expression of linc-MAF-4 and MAF assessed at different time points by RT-qPCR in activated CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions (average of four technical replicates ± SEM). See also Supplementary Fig. 4 b,c . ( b ) ChIP-qPCR analysis of H3K4me3 and RNA polymerase II occupancy at MAF locus in CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions at day 8 post-activation. Enrichment is a percentage of input (average of at least 5 independent experiments ± SEM). One-tailed t -test * P

    Journal: Nature immunology

    Article Title: LincRNA landscape in human lymphocytes highlights regulation of T cell differentiation by linc-MAF-4

    doi: 10.1038/ni.3093

    Figure Lengend Snippet: Linc-MAF-4 contributes to T H 1 cell differentiation ( a ) Expression of linc-MAF-4 and MAF assessed at different time points by RT-qPCR in activated CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions (average of four technical replicates ± SEM). See also Supplementary Fig. 4 b,c . ( b ) ChIP-qPCR analysis of H3K4me3 and RNA polymerase II occupancy at MAF locus in CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions at day 8 post-activation. Enrichment is a percentage of input (average of at least 5 independent experiments ± SEM). One-tailed t -test * P

    Article Snippet: Indeed, linc-MAF-4, one of the TH 1 signature lincRNA, was poorly expressed in TH 2 cells and its experimental downregulation skewed differentiating T helper cells toward a TH 2 transcription profile.

    Techniques: Cell Differentiation, Expressing, Quantitative RT-PCR, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Activation Assay, One-tailed Test

    Epigenetic characterization of linc-MAF4/MAF genomic locus ( a ) Schematic representation of the genomic region analyzed by 3C. Position relative to linc-MAF-4 and MAF of three lincRNAs present in the region is shown in the upper part of the panel. The M1 primer at the 5′ end of MAF (red line) was used as bait. Primers (L1-L24) spanning the region between linc-MAF-4 and MAF were tested for interaction. Relative frequency of interaction between MAF and linc-MAF-4 5 ′ (L7) and 3 ′ (L12) ends is shown in CD4 + naïve T cells differentiated in T H 1 polarizing conditions (day 8) (average of three independent experiments ± SEM). ( b ) Relative abundance of linc-MAF-4 transcript in cytoplasm, nucleus and chromatin in CD4 + naïve T cells differentiated in T H 1 polarizing conditions (day 8). Linc-00339, MALAT1 and RNU2.1 were used respectively as cytoplasmic, nuclear and chromatin-associated controls (average of three independent experiments ± SEM). ( c ) RIP assay for LSD1 and EZH2 in CD4 + naïve T cells differentiated in T H 1 polarizing conditions (day 8). Fold enrichment is relative to mock. ACTB, RNU2.1 and a region upstream the TSS of linc-MAF-4 were chosen as controls (average of six independent experiments ± SEM). The statistical significance was determined with ANOVA and Dunnet post-hoc test: * P

    Journal: Nature immunology

    Article Title: LincRNA landscape in human lymphocytes highlights regulation of T cell differentiation by linc-MAF-4

    doi: 10.1038/ni.3093

    Figure Lengend Snippet: Epigenetic characterization of linc-MAF4/MAF genomic locus ( a ) Schematic representation of the genomic region analyzed by 3C. Position relative to linc-MAF-4 and MAF of three lincRNAs present in the region is shown in the upper part of the panel. The M1 primer at the 5′ end of MAF (red line) was used as bait. Primers (L1-L24) spanning the region between linc-MAF-4 and MAF were tested for interaction. Relative frequency of interaction between MAF and linc-MAF-4 5 ′ (L7) and 3 ′ (L12) ends is shown in CD4 + naïve T cells differentiated in T H 1 polarizing conditions (day 8) (average of three independent experiments ± SEM). ( b ) Relative abundance of linc-MAF-4 transcript in cytoplasm, nucleus and chromatin in CD4 + naïve T cells differentiated in T H 1 polarizing conditions (day 8). Linc-00339, MALAT1 and RNU2.1 were used respectively as cytoplasmic, nuclear and chromatin-associated controls (average of three independent experiments ± SEM). ( c ) RIP assay for LSD1 and EZH2 in CD4 + naïve T cells differentiated in T H 1 polarizing conditions (day 8). Fold enrichment is relative to mock. ACTB, RNU2.1 and a region upstream the TSS of linc-MAF-4 were chosen as controls (average of six independent experiments ± SEM). The statistical significance was determined with ANOVA and Dunnet post-hoc test: * P

    Article Snippet: Indeed, linc-MAF-4, one of the TH 1 signature lincRNA, was poorly expressed in TH 2 cells and its experimental downregulation skewed differentiating T helper cells toward a TH 2 transcription profile.

    Techniques:

    Power for the W2WK (red), traditional joint (blue), and traditional main effect (green) approaches by causal SNP under the M1 model. An “x” marks the 14 SNPs that were modeled as genotyped in our simulations. The MAF of each SNP (grey line) is along the right Y-axis. Plot on the left assumes an additive model; the plot on the right assumes the underlying model is dominant, but was tested as additive. Inset plot shows the underlying model (M1): solid black line represents genotypic effect among individuals who have been exposed to the environmental insult. Dotted black line shows genotypic effect of individuals who were not exposed to the environmental insult under the M1 model. Dotted grey lines indicate alternate models that are considered elsewhere in this manuscript.

    Journal: Genetic epidemiology

    Article Title: Kernel Approach for Modeling Interaction Effects in Genetic Association Studies of Complex Quantitative Traits

    doi: 10.1002/gepi.21901

    Figure Lengend Snippet: Power for the W2WK (red), traditional joint (blue), and traditional main effect (green) approaches by causal SNP under the M1 model. An “x” marks the 14 SNPs that were modeled as genotyped in our simulations. The MAF of each SNP (grey line) is along the right Y-axis. Plot on the left assumes an additive model; the plot on the right assumes the underlying model is dominant, but was tested as additive. Inset plot shows the underlying model (M1): solid black line represents genotypic effect among individuals who have been exposed to the environmental insult. Dotted black line shows genotypic effect of individuals who were not exposed to the environmental insult under the M1 model. Dotted grey lines indicate alternate models that are considered elsewhere in this manuscript.

    Article Snippet: For example, SNP 9 (rs9909659) has a MAF 0.21 and is not genotyped on the Illumina array.

    Techniques:

    Power for the W2WK (red), traditional joint (blue), and traditional main effect (green) approaches by causal SNP under the M5 model. An “x” marks the 14 SNPs that were modeled as genotyped in our simulations. The MAF of each SNP (grey line) is along the right Y-axis. Plot on the left assumes an additive model; the plot on the right assumes the underlying model is dominant, but was tested as additive. Inset plot shows the underlying model (M5): solid black line represents genotypic effect among individuals who have been exposed to the environmental insult. Dotted black line shows genotypic effect of individuals who were not exposed to the environmental insult under the M5 model. Dotted grey lines indicate alternate models that are considered elsewhere in this manuscript.

    Journal: Genetic epidemiology

    Article Title: Kernel Approach for Modeling Interaction Effects in Genetic Association Studies of Complex Quantitative Traits

    doi: 10.1002/gepi.21901

    Figure Lengend Snippet: Power for the W2WK (red), traditional joint (blue), and traditional main effect (green) approaches by causal SNP under the M5 model. An “x” marks the 14 SNPs that were modeled as genotyped in our simulations. The MAF of each SNP (grey line) is along the right Y-axis. Plot on the left assumes an additive model; the plot on the right assumes the underlying model is dominant, but was tested as additive. Inset plot shows the underlying model (M5): solid black line represents genotypic effect among individuals who have been exposed to the environmental insult. Dotted black line shows genotypic effect of individuals who were not exposed to the environmental insult under the M5 model. Dotted grey lines indicate alternate models that are considered elsewhere in this manuscript.

    Article Snippet: For example, SNP 9 (rs9909659) has a MAF 0.21 and is not genotyped on the Illumina array.

    Techniques:

    Sox5 and c-Maf activate the promoter of RORγt gene. (A, top) VISTA plot of CNSs of Rorc gene locus. (middle panels) Naive CD4 + T cells from CD4 cre Sox5 fl/fl or Sox5 fl/fl mice were stimulated under Th17 conditions for 8 h. ChIP-qPCR assay for CNS of Rorc gene locus was performed with anti–c-Maf antibody, anti-p300 antibody, or control rabbit IgG. (bottom) Naive CD4 + T cells from WT mice were stimulated under Th17 conditions for 8 h. ChIP-qPCR assay for CNS of Rorc gene locus was performed with anti-Stat3 antibody, anti-H3K4me3 antibody, or control rabbit IgG. Results are expressed as the percent input for each ChIP fraction. Data are representative of three independent experiments. (B) Luciferase assay of indicated RORγt-Luci in EL4 cells. Data are compiled from three independent experiments. (C) Sequence of RORγt promoter (from −1648 to −1448). Putative Sox5 binding site is indicated by red and putative MAREs are indicated by blue. Mutated sequences are shown in lower lines. (D, top) Representative data of DNA precipitation assay of HA-Sox5 with biotinylated double-stranded DNA probes containing a portion of RORγt promoter or its mutant at Sox5-binding site. As a positive control (PC), FXO+ probe, which harbors two Sox consensus sites, was used. (bottom) Representative data of DNA-precipitation assay of c-Maf with biotinylated double-stranded DNA probes containing a portion of RORγt promoter or its mutants at MARE sites. Shown are data representative of three independent experiments. (E) Naive CD4 + T cells were co-infected with retroviruses of either pMX-IN or pMX-IN-Sox5t and with those of either MIT or MIT-c-Maf for 24 h. Cells were reinfected with WT-RORγtP for 24 h, and GFP versus hCD4 in NGFR + Thy1.1 + cells was evaluated by flow cytometry. (F) Naive CD4 + T cells were co-infected with retroviruses of indicated vectors and analyzed as described in Fig. 7 E . (G) Naive CD4 + T cells from Sox5 fl/fl mice and CD4 cre Sox5 fl/fl mice were stimulated in neutral conditions for 48 h and infected with retroviruses of hCD4-pA-GFP (empty), WT-RORγtP, or ΔSox5-ΔMARE-RORγtP under Th17 conditions for additional 24 h. The expression levels of GFP were evaluated as described in Fig. 7 E . Data shown are representative of three independent experiments.

    Journal: The Journal of Experimental Medicine

    Article Title: Sox5 and c-Maf cooperatively induce Th17 cell differentiation via RORγt induction as downstream targets of Stat3

    doi: 10.1084/jem.20130791

    Figure Lengend Snippet: Sox5 and c-Maf activate the promoter of RORγt gene. (A, top) VISTA plot of CNSs of Rorc gene locus. (middle panels) Naive CD4 + T cells from CD4 cre Sox5 fl/fl or Sox5 fl/fl mice were stimulated under Th17 conditions for 8 h. ChIP-qPCR assay for CNS of Rorc gene locus was performed with anti–c-Maf antibody, anti-p300 antibody, or control rabbit IgG. (bottom) Naive CD4 + T cells from WT mice were stimulated under Th17 conditions for 8 h. ChIP-qPCR assay for CNS of Rorc gene locus was performed with anti-Stat3 antibody, anti-H3K4me3 antibody, or control rabbit IgG. Results are expressed as the percent input for each ChIP fraction. Data are representative of three independent experiments. (B) Luciferase assay of indicated RORγt-Luci in EL4 cells. Data are compiled from three independent experiments. (C) Sequence of RORγt promoter (from −1648 to −1448). Putative Sox5 binding site is indicated by red and putative MAREs are indicated by blue. Mutated sequences are shown in lower lines. (D, top) Representative data of DNA precipitation assay of HA-Sox5 with biotinylated double-stranded DNA probes containing a portion of RORγt promoter or its mutant at Sox5-binding site. As a positive control (PC), FXO+ probe, which harbors two Sox consensus sites, was used. (bottom) Representative data of DNA-precipitation assay of c-Maf with biotinylated double-stranded DNA probes containing a portion of RORγt promoter or its mutants at MARE sites. Shown are data representative of three independent experiments. (E) Naive CD4 + T cells were co-infected with retroviruses of either pMX-IN or pMX-IN-Sox5t and with those of either MIT or MIT-c-Maf for 24 h. Cells were reinfected with WT-RORγtP for 24 h, and GFP versus hCD4 in NGFR + Thy1.1 + cells was evaluated by flow cytometry. (F) Naive CD4 + T cells were co-infected with retroviruses of indicated vectors and analyzed as described in Fig. 7 E . (G) Naive CD4 + T cells from Sox5 fl/fl mice and CD4 cre Sox5 fl/fl mice were stimulated in neutral conditions for 48 h and infected with retroviruses of hCD4-pA-GFP (empty), WT-RORγtP, or ΔSox5-ΔMARE-RORγtP under Th17 conditions for additional 24 h. The expression levels of GFP were evaluated as described in Fig. 7 E . Data shown are representative of three independent experiments.

    Article Snippet: The following antibodies were used for ChIP: anti-c-Maf antibody (M153; Santa Cruz Biotechnology, Inc.), anti-HA antibody (ab9110; Abcam), anti-p300 antibody (C20; Santa Cruz Biotechnology, Inc.), and anti-H3K4me3 antibody (17–614; Millipore).

    Techniques: Mouse Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Luciferase, Sequencing, Binding Assay, Mutagenesis, Positive Control, Infection, Flow Cytometry, Cytometry, Expressing

    LiCl-mediated GSK3 inhibition leads to MAFB and c-MAF degradation and decreases DEPTOR expression. RPMI ( a – c ) and L363 ( b – d ) cells were treated or not with LiCl for 1 h and then with MG132 (MG) ( a and b ) or bortezomib (Btz) ( c – d ) for 3 h. Anti-c-MAF, MAFB, p-MAF and SAM68 western blots were carried out on nuclear extracts. RPMI ( e ) and L363 ( f ) cells were treated or not treated (w/o) with LiCl followed by treatment with 20 μg/ml cycloheximide (Chx) for the indicated time points. Nuclear extracts were analysed by western blots using c-MAF, MAFB, p-Maf and SAM68 antibodies. Band intensities were measured, normalised to SAM68 and plotted as a percentage of the initial band intensity (right panels). ( g ) The Relative DEPTOR expression in different HMCLs treated with LiCl (as indicated) was determined by RT-qPCR. Expression was normalised to HPRT. NS, non-specific; P, phosphorylated forms. DEPTOR expression was specifically decreased upon LiCl treatment in MAFB (L363 P=0.07, OPM2 P=NS) and c-MAF-expressing cells (RPMI P

    Journal: Blood Cancer Journal

    Article Title: GSK3-mediated MAF phosphorylation in multiple myeloma as a potential therapeutic target

    doi: 10.1038/bcj.2013.67

    Figure Lengend Snippet: LiCl-mediated GSK3 inhibition leads to MAFB and c-MAF degradation and decreases DEPTOR expression. RPMI ( a – c ) and L363 ( b – d ) cells were treated or not with LiCl for 1 h and then with MG132 (MG) ( a and b ) or bortezomib (Btz) ( c – d ) for 3 h. Anti-c-MAF, MAFB, p-MAF and SAM68 western blots were carried out on nuclear extracts. RPMI ( e ) and L363 ( f ) cells were treated or not treated (w/o) with LiCl followed by treatment with 20 μg/ml cycloheximide (Chx) for the indicated time points. Nuclear extracts were analysed by western blots using c-MAF, MAFB, p-Maf and SAM68 antibodies. Band intensities were measured, normalised to SAM68 and plotted as a percentage of the initial band intensity (right panels). ( g ) The Relative DEPTOR expression in different HMCLs treated with LiCl (as indicated) was determined by RT-qPCR. Expression was normalised to HPRT. NS, non-specific; P, phosphorylated forms. DEPTOR expression was specifically decreased upon LiCl treatment in MAFB (L363 P=0.07, OPM2 P=NS) and c-MAF-expressing cells (RPMI P

    Article Snippet: Protein extracts were subjected to SDS-PAGE, transferred to Immobilon-P membranes (Millipore, Molsheim, France) and probed with anti-MafA (Abcam, Paris, France, 1/2000), anti-MAFB (Santa Cruz, Heidelberg, Germany, sc-10022 p20, 1/2000), anti-c-MAF (Santa Cruz, M153, 1/2000), anti-SAM68 (Santa Cruz, 1/5000), anti-β-actin (Sigma, 1/50000) and rabbit polyclonal T53-T57 phospho-specific (Rocques et al. , 1/2000) antibodies.

    Techniques: Inhibition, Expressing, Western Blot, Quantitative RT-PCR

    MAFB and c-MAF are phosphorylated by GSK3 in MM. Relative c-MAF ( a ) and MAFB ( b ) expression in MM cell lines (HMCLs) was determined by RT-qPCR. HMCL's nuclear extracts were analysed by western blot using c-MAF ( c ) and MAFB ( d ) antibodies, p-MAF and SAM68 antibodies. c-MAF ( e ) and MAFB ( f ) expressing cell lines were treated with LiCl as indicated. Anti-c-MAF, anti-MAFB, p-MAF and SAM68 western blots were carried out on nuclear extracts. NS, non-specific; P, phosphorylated forms.

    Journal: Blood Cancer Journal

    Article Title: GSK3-mediated MAF phosphorylation in multiple myeloma as a potential therapeutic target

    doi: 10.1038/bcj.2013.67

    Figure Lengend Snippet: MAFB and c-MAF are phosphorylated by GSK3 in MM. Relative c-MAF ( a ) and MAFB ( b ) expression in MM cell lines (HMCLs) was determined by RT-qPCR. HMCL's nuclear extracts were analysed by western blot using c-MAF ( c ) and MAFB ( d ) antibodies, p-MAF and SAM68 antibodies. c-MAF ( e ) and MAFB ( f ) expressing cell lines were treated with LiCl as indicated. Anti-c-MAF, anti-MAFB, p-MAF and SAM68 western blots were carried out on nuclear extracts. NS, non-specific; P, phosphorylated forms.

    Article Snippet: Protein extracts were subjected to SDS-PAGE, transferred to Immobilon-P membranes (Millipore, Molsheim, France) and probed with anti-MafA (Abcam, Paris, France, 1/2000), anti-MAFB (Santa Cruz, Heidelberg, Germany, sc-10022 p20, 1/2000), anti-c-MAF (Santa Cruz, M153, 1/2000), anti-SAM68 (Santa Cruz, 1/5000), anti-β-actin (Sigma, 1/50000) and rabbit polyclonal T53-T57 phospho-specific (Rocques et al. , 1/2000) antibodies.

    Techniques: Expressing, Quantitative RT-PCR, Western Blot

    Highly efficient target gene KO in nonactivated human and mouse primary T cells with an optimized CRISPR/Cas9 RNP transfection approach. (A–D) KO efficiency by flow cytometry of CXCR4, CD127, and CCR7 in nonactivated human CD4 T cells 72 h after transfection (A); PD1, TIGIT, and CTLA4 in nonactivated human CD8 T cells 72 h after transfection followed by 72 h of stimulation with anti-CD3/anti-CD28 (B); CD90 and CTLA4 in IL-7–preconditioned nonactivated mouse CD4 T cells 5 d after transfection and incubation with IL-7 (C); and CD8α and CTLA4 in IL-7–preconditioned nonactivated mouse CD8 + T cells after transfection and 5 d of incubation in the presence of IL-7 followed by 48 h stimulation with anti-CD3/anti-CD28 (D; all compared with nontargeting control Cas9 RNP-transfected T cells). Data are presented as mean ± SD ( n = 2) and representative of three (A) or two (B–D) independent experiments. (E) IFN-γ expression by flow cytometry in crIFNγ or NTC-transfected nonactivated human CD8 + T cells cultured for 3 d and restimulated for 4 h with PMA/ionomycin. Data are presented as mean ± SD ( n = 2) and representative of two independent experiments. (F) FoxP3 and c-Maf expression by flow cytometry in crFoxp3 or NTC transfected IL-7–preconditioned nonactivated mouse CD4 + T cells 5 d after transfection with IL-7 followed by 3 d of polarization into iTreg with TGF-β and IL-2. Data are presented as mean ± SD ( n = 2) and representative of two independent experiments. (G–I) KO efficiency by flow cytometry of double KOs targeting CD127 and CCR7 in nonactivated human CD4 + T cells 72 h after transfection (G), CD90 and CTLA4 in nonactivated mouse CD8 T cells after transfection and 5 d of incubation in the presence of IL-7 followed by 48 h stimulation with anti-CD3/anti-CD28 (H), or CD90 and FoxP3 in nonactivated mouse CD4 + T cells after transfection and 5 d of incubation with IL-7 followed by 3 d of polarization into iTreg with TGF-β and IL-2 with anti-CD3/anti-CD28 (I). Data are presented as mean ± SD ( n = 2) and representative of two independent experiments. ***, P

    Journal: The Journal of Experimental Medicine

    Article Title: Optimized RNP transfection for highly efficient CRISPR/Cas9-mediated gene knockout in primary T cells

    doi: 10.1084/jem.20171626

    Figure Lengend Snippet: Highly efficient target gene KO in nonactivated human and mouse primary T cells with an optimized CRISPR/Cas9 RNP transfection approach. (A–D) KO efficiency by flow cytometry of CXCR4, CD127, and CCR7 in nonactivated human CD4 T cells 72 h after transfection (A); PD1, TIGIT, and CTLA4 in nonactivated human CD8 T cells 72 h after transfection followed by 72 h of stimulation with anti-CD3/anti-CD28 (B); CD90 and CTLA4 in IL-7–preconditioned nonactivated mouse CD4 T cells 5 d after transfection and incubation with IL-7 (C); and CD8α and CTLA4 in IL-7–preconditioned nonactivated mouse CD8 + T cells after transfection and 5 d of incubation in the presence of IL-7 followed by 48 h stimulation with anti-CD3/anti-CD28 (D; all compared with nontargeting control Cas9 RNP-transfected T cells). Data are presented as mean ± SD ( n = 2) and representative of three (A) or two (B–D) independent experiments. (E) IFN-γ expression by flow cytometry in crIFNγ or NTC-transfected nonactivated human CD8 + T cells cultured for 3 d and restimulated for 4 h with PMA/ionomycin. Data are presented as mean ± SD ( n = 2) and representative of two independent experiments. (F) FoxP3 and c-Maf expression by flow cytometry in crFoxp3 or NTC transfected IL-7–preconditioned nonactivated mouse CD4 + T cells 5 d after transfection with IL-7 followed by 3 d of polarization into iTreg with TGF-β and IL-2. Data are presented as mean ± SD ( n = 2) and representative of two independent experiments. (G–I) KO efficiency by flow cytometry of double KOs targeting CD127 and CCR7 in nonactivated human CD4 + T cells 72 h after transfection (G), CD90 and CTLA4 in nonactivated mouse CD8 T cells after transfection and 5 d of incubation in the presence of IL-7 followed by 48 h stimulation with anti-CD3/anti-CD28 (H), or CD90 and FoxP3 in nonactivated mouse CD4 + T cells after transfection and 5 d of incubation with IL-7 followed by 3 d of polarization into iTreg with TGF-β and IL-2 with anti-CD3/anti-CD28 (I). Data are presented as mean ± SD ( n = 2) and representative of two independent experiments. ***, P

    Article Snippet: For mouse cells, CD4 (RM4-5; eBioscience), CD8a (53-6.7; BD Biosciences), CD90.2 (53-2.1, eBioscience), PD-1 (29F.1A12; BioLegend), CTLA-4 (UC10-4B9, eBioscience), CD25 (PC61; BD Biosciences), CD69 (H1.2F3; BD Biosciences), Foxp3 (FJK-16s; eBioscience), and c-Maf (sym0F1, eBioscience) were used.

    Techniques: CRISPR, Transfection, Flow Cytometry, Cytometry, Incubation, Expressing, Cell Culture

    Expression of c-Maf and its target genes in the dorsal horn of MafA −/− mice. A , B , Immunostaining of c-Maf was performed on sections of spinal cord in MafA +/− and MafA −/− mice at P0. Insets show the anti-c-Maf immunofluorescence signal in the DRG of control and MafA deletion mice. Note comparable expression of c-Maf in both the dorsal horn and the DRG in MafA −/− versus control mice. C–H , In situ hybridization was performed with Gabra5 ( C , D ), CCK ( E , F ), and Rora ( G , H ) probes on sections of cervical/thoracic spinal cord in MafA +/− and MafA −/− mice at P0. No apparent difference was observed in MafA mutant and control mice. Scale bars: (in B , H ) A–H , 50 μm; (in B , inset), A , B , insets, 50 μm.

    Journal: The Journal of Neuroscience

    Article Title: c-Maf Is Required for the Development of Dorsal Horn Laminae III/IV Neurons and Mechanoreceptive DRG Axon Projections

    doi: 10.1523/JNEUROSCI.6239-11.2012

    Figure Lengend Snippet: Expression of c-Maf and its target genes in the dorsal horn of MafA −/− mice. A , B , Immunostaining of c-Maf was performed on sections of spinal cord in MafA +/− and MafA −/− mice at P0. Insets show the anti-c-Maf immunofluorescence signal in the DRG of control and MafA deletion mice. Note comparable expression of c-Maf in both the dorsal horn and the DRG in MafA −/− versus control mice. C–H , In situ hybridization was performed with Gabra5 ( C , D ), CCK ( E , F ), and Rora ( G , H ) probes on sections of cervical/thoracic spinal cord in MafA +/− and MafA −/− mice at P0. No apparent difference was observed in MafA mutant and control mice. Scale bars: (in B , H ) A–H , 50 μm; (in B , inset), A , B , insets, 50 μm.

    Article Snippet: The following antibodies were used: rabbit anti-c-Maf (1:500, Bethyl Laboratories); rabbit anti-MafA (1:50, Bethyl Laboratories); rabbit anti-Pax2 (1:50, Zymed); mouse anti-NF200 (neurofilament-200) (1:1000, Sigma); mouse anti-NeuN (1:1000, Millipore); rabbit anti-S100 (1:400, Dako); rabbit anti-TrkA (1:100, Advanced Targeting Systems); guinea-pig anti-VGLUT1 (1:100, Millipore); rabbit anti-parvalbumin (1:100, Swant); goat anti-Ret (1:25, R & D Systems); and chicken anti-β-gal (1:250, Abcam).

    Techniques: Expressing, Mouse Assay, Immunostaining, Immunofluorescence, In Situ Hybridization, Mutagenesis

    Linc-MAF-4 contributes to T H 1 cell differentiation ( a ) Expression of linc-MAF-4 and MAF assessed at different time points by RT-qPCR in activated CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions (average of four technical replicates ± SEM). See also Supplementary Fig. 4 b,c . ( b ) ChIP-qPCR analysis of H3K4me3 and RNA polymerase II occupancy at MAF locus in CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions at day 8 post-activation. Enrichment is a percentage of input (average of at least 5 independent experiments ± SEM). One-tailed t -test * P

    Journal: Nature immunology

    Article Title: LincRNA landscape in human lymphocytes highlights regulation of T cell differentiation by linc-MAF-4

    doi: 10.1038/ni.3093

    Figure Lengend Snippet: Linc-MAF-4 contributes to T H 1 cell differentiation ( a ) Expression of linc-MAF-4 and MAF assessed at different time points by RT-qPCR in activated CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions (average of four technical replicates ± SEM). See also Supplementary Fig. 4 b,c . ( b ) ChIP-qPCR analysis of H3K4me3 and RNA polymerase II occupancy at MAF locus in CD4 + naïve T cells differentiated in T H 1 or T H 2 polarizing conditions at day 8 post-activation. Enrichment is a percentage of input (average of at least 5 independent experiments ± SEM). One-tailed t -test * P

    Article Snippet: Diluted cDNA was then used as input for RT-qPCR to assess MAF (Hs00193519_m1), IL4 (Hs00174122_m1), GATA3 (Hs01651755_m1), TBX21 (Hs00203436_m1), RORC (Hs01076119_m1), IL17 (Hs00174383_m1), Linc00339 (Hs04331223_m1), MALAT1 (Hs01910177_s1), RNU2.1 (Hs03023892_g1) and GAPDH (Hs02758991_g1) gene expression with Inventoried TaqMan Gene Expression assays (LifeTechnologies) were used.

    Techniques: Cell Differentiation, Expressing, Quantitative RT-PCR, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Activation Assay, One-tailed Test

    Epigenetic characterization of linc-MAF4/MAF genomic locus ( a ) Schematic representation of the genomic region analyzed by 3C. Position relative to linc-MAF-4 and MAF of three lincRNAs present in the region is shown in the upper part of the panel. The M1 primer at the 5′ end of MAF (red line) was used as bait. Primers (L1-L24) spanning the region between linc-MAF-4 and MAF were tested for interaction. Relative frequency of interaction between MAF and linc-MAF-4 5 ′ (L7) and 3 ′ (L12) ends is shown in CD4 + naïve T cells differentiated in T H 1 polarizing conditions (day 8) (average of three independent experiments ± SEM). ( b ) Relative abundance of linc-MAF-4 transcript in cytoplasm, nucleus and chromatin in CD4 + naïve T cells differentiated in T H 1 polarizing conditions (day 8). Linc-00339, MALAT1 and RNU2.1 were used respectively as cytoplasmic, nuclear and chromatin-associated controls (average of three independent experiments ± SEM). ( c ) RIP assay for LSD1 and EZH2 in CD4 + naïve T cells differentiated in T H 1 polarizing conditions (day 8). Fold enrichment is relative to mock. ACTB, RNU2.1 and a region upstream the TSS of linc-MAF-4 were chosen as controls (average of six independent experiments ± SEM). The statistical significance was determined with ANOVA and Dunnet post-hoc test: * P

    Journal: Nature immunology

    Article Title: LincRNA landscape in human lymphocytes highlights regulation of T cell differentiation by linc-MAF-4

    doi: 10.1038/ni.3093

    Figure Lengend Snippet: Epigenetic characterization of linc-MAF4/MAF genomic locus ( a ) Schematic representation of the genomic region analyzed by 3C. Position relative to linc-MAF-4 and MAF of three lincRNAs present in the region is shown in the upper part of the panel. The M1 primer at the 5′ end of MAF (red line) was used as bait. Primers (L1-L24) spanning the region between linc-MAF-4 and MAF were tested for interaction. Relative frequency of interaction between MAF and linc-MAF-4 5 ′ (L7) and 3 ′ (L12) ends is shown in CD4 + naïve T cells differentiated in T H 1 polarizing conditions (day 8) (average of three independent experiments ± SEM). ( b ) Relative abundance of linc-MAF-4 transcript in cytoplasm, nucleus and chromatin in CD4 + naïve T cells differentiated in T H 1 polarizing conditions (day 8). Linc-00339, MALAT1 and RNU2.1 were used respectively as cytoplasmic, nuclear and chromatin-associated controls (average of three independent experiments ± SEM). ( c ) RIP assay for LSD1 and EZH2 in CD4 + naïve T cells differentiated in T H 1 polarizing conditions (day 8). Fold enrichment is relative to mock. ACTB, RNU2.1 and a region upstream the TSS of linc-MAF-4 were chosen as controls (average of six independent experiments ± SEM). The statistical significance was determined with ANOVA and Dunnet post-hoc test: * P

    Article Snippet: Diluted cDNA was then used as input for RT-qPCR to assess MAF (Hs00193519_m1), IL4 (Hs00174122_m1), GATA3 (Hs01651755_m1), TBX21 (Hs00203436_m1), RORC (Hs01076119_m1), IL17 (Hs00174383_m1), Linc00339 (Hs04331223_m1), MALAT1 (Hs01910177_s1), RNU2.1 (Hs03023892_g1) and GAPDH (Hs02758991_g1) gene expression with Inventoried TaqMan Gene Expression assays (LifeTechnologies) were used.

    Techniques:

    Cholesterol biosynthesis pathway inhibition interferes with c-Maf expression. Purified human CD4 + T cells stimulated in vitro with plate-bound α-CD3 (2 μgml −1 ) + α-CD46 (5 μgml −1 ) and recombinant human interleukin-2 (rhIL-2) (50 Uml −1 ) were cultured for 36 h in the presence of 25-hydroxycholesterol (25-HC). a Expression levels of MAF mRNA ( n = 7). b Expression levels of PRDM1 mRNA ( n = 6). c Representative flow cytometric analysis of intracellular c-Maf staining. d Normalised frequency of c-Maf + cells ( n = 16). e Normalised frequency of c-Maf + cells cultured under fully supplemented medium (M) and cholesterol (500×) (Ch) in the presence or absence of 2μM 25-HC ( n = 3). f Correlation between frequency of c-Maf + and IL-10 + cells, numbers denote r and p values for Spearman's correlation test. Graphs show independent donors (dots) normalised to untreated cells; bars represent median values. *

    Journal: Nature Communications

    Article Title: The cholesterol biosynthesis pathway regulates IL-10 expression in human Th1 cells

    doi: 10.1038/s41467-019-08332-9

    Figure Lengend Snippet: Cholesterol biosynthesis pathway inhibition interferes with c-Maf expression. Purified human CD4 + T cells stimulated in vitro with plate-bound α-CD3 (2 μgml −1 ) + α-CD46 (5 μgml −1 ) and recombinant human interleukin-2 (rhIL-2) (50 Uml −1 ) were cultured for 36 h in the presence of 25-hydroxycholesterol (25-HC). a Expression levels of MAF mRNA ( n = 7). b Expression levels of PRDM1 mRNA ( n = 6). c Representative flow cytometric analysis of intracellular c-Maf staining. d Normalised frequency of c-Maf + cells ( n = 16). e Normalised frequency of c-Maf + cells cultured under fully supplemented medium (M) and cholesterol (500×) (Ch) in the presence or absence of 2μM 25-HC ( n = 3). f Correlation between frequency of c-Maf + and IL-10 + cells, numbers denote r and p values for Spearman's correlation test. Graphs show independent donors (dots) normalised to untreated cells; bars represent median values. *

    Article Snippet: Normalised RNA quantities were reverse transcribed into cDNA using the qPCRBIO Synthesis kit (PCR Biosystems). qPCR was performed in qPCRBIO Probe Mix (PCR Biosystems) using pre-validated FAM-labelled Taqman Gene Expression Assays for LDLr (Hs00181192_m1), HMGCS1 (Hs00940429_m1), FDFT1 (Hs00926054_m1) and DHCR7 (Hs01023087_m1), IL10 (Hs00961622_m1), MAF (Hs04185012_s1) and PRDM1 (Hs00153357_m1).

    Techniques: Inhibition, Expressing, Purification, In Vitro, Recombinant, Cell Culture, Flow Cytometry, Staining

    Messenger RNA (mRNA) levels of cholesterol biosynthesis pathway (CBP) enzymes are related to disease progression in man. a Expression levels of cholesterol-25-hydroxylase ( CH25H ), FDFT1 and DHCR7 obtained from a gene expression profiling study of synovial biopsies from 13 rheumatoid arthritis (RA)-risk individuals who were followed over time to investigate the development of arthritis. RA-risk individuals were stratified into relative low or high expressers for the indicated genes, after which their arthritis-free survival was compared using a log-rank (Mantel–Cox) test. Graphs display arthritis-free survival curves including the 95% confidence interval (CI). b Schematic representation of interleukin-10 (IL-10) regulation by the cholesterol biosynthesis pathway. Normal cholesterol pathway activity is required for adequate transcription of the IL10 gene and IL-10 expression in T-helper effector cells (left); when the flux is reduced via statin or 25-hydroxycholesterol (25-HC) supplementation, c-Maf expression is downregulated and IL-10 gene transcription reduced

    Journal: Nature Communications

    Article Title: The cholesterol biosynthesis pathway regulates IL-10 expression in human Th1 cells

    doi: 10.1038/s41467-019-08332-9

    Figure Lengend Snippet: Messenger RNA (mRNA) levels of cholesterol biosynthesis pathway (CBP) enzymes are related to disease progression in man. a Expression levels of cholesterol-25-hydroxylase ( CH25H ), FDFT1 and DHCR7 obtained from a gene expression profiling study of synovial biopsies from 13 rheumatoid arthritis (RA)-risk individuals who were followed over time to investigate the development of arthritis. RA-risk individuals were stratified into relative low or high expressers for the indicated genes, after which their arthritis-free survival was compared using a log-rank (Mantel–Cox) test. Graphs display arthritis-free survival curves including the 95% confidence interval (CI). b Schematic representation of interleukin-10 (IL-10) regulation by the cholesterol biosynthesis pathway. Normal cholesterol pathway activity is required for adequate transcription of the IL10 gene and IL-10 expression in T-helper effector cells (left); when the flux is reduced via statin or 25-hydroxycholesterol (25-HC) supplementation, c-Maf expression is downregulated and IL-10 gene transcription reduced

    Article Snippet: Normalised RNA quantities were reverse transcribed into cDNA using the qPCRBIO Synthesis kit (PCR Biosystems). qPCR was performed in qPCRBIO Probe Mix (PCR Biosystems) using pre-validated FAM-labelled Taqman Gene Expression Assays for LDLr (Hs00181192_m1), HMGCS1 (Hs00940429_m1), FDFT1 (Hs00926054_m1) and DHCR7 (Hs01023087_m1), IL10 (Hs00961622_m1), MAF (Hs04185012_s1) and PRDM1 (Hs00153357_m1).

    Techniques: Expressing, Activity Assay