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Pyrosequencing Inc tsdr pyrosequencing primer agaaatttgtggggtggg
TYK2 inhibition enhances Treg stability in Th17-polarizing conditions Tregs (CD4 + CD25 hi CD127 lo ) were isolated from peripheral blood of healthy human donors, stimulated with anti-CD3/CD28, and cultured for 7 days in a Th17-polarizing cocktail in the presence of BMS-986202 (B202). (A) Schematic of polarization protocol. (B) The proportion of FOXP3 + cells was assessed after 7 days. Representative plots and quantification are shown ( n = 9). (C) Expression of RORC2, CCR4, and CCR6 was determined after 7 days relative to non-polarized, control Tregs ( n = 3). (D) Methylation of the Treg-specific demethylated region <t>(TSDR)</t> after 7 days of culture ( n = 5). (E) Representative plot and quantification of intracellular IL-17A/F expression after a 4 h stimulation with PMA/ionomycin ( n = 5). (F) IL-17A, IL-17F, and TNF-α concentration in supernatant after a 24 h re-stimulation with anti-CD3/CD28 ( n = 6). (G) Representative figures and quantification of TIGIT and CD226 expression on Tregs on day 7 ( n = 9). Statistically significant differences compared to DMSO-treated cells were determined by a repeated-measures one-way ANOVA with Dunnett’s multiple comparisons test (B–G). Bars indicate mean ± SEM. Points indicate individual replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.
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TYK2 inhibition enhances Treg stability in Th17-polarizing conditions Tregs (CD4 + CD25 hi CD127 lo ) were isolated from peripheral blood of healthy human donors, stimulated with anti-CD3/CD28, and cultured for 7 days in a Th17-polarizing cocktail in the presence of BMS-986202 (B202). (A) Schematic of polarization protocol. (B) The proportion of FOXP3 + cells was assessed after 7 days. Representative plots and quantification are shown ( n = 9). (C) Expression of RORC2, CCR4, and CCR6 was determined after 7 days relative to non-polarized, control Tregs ( n = 3). (D) Methylation of the Treg-specific demethylated region <t>(TSDR)</t> after 7 days of culture ( n = 5). (E) Representative plot and quantification of intracellular IL-17A/F expression after a 4 h stimulation with PMA/ionomycin ( n = 5). (F) IL-17A, IL-17F, and TNF-α concentration in supernatant after a 24 h re-stimulation with anti-CD3/CD28 ( n = 6). (G) Representative figures and quantification of TIGIT and CD226 expression on Tregs on day 7 ( n = 9). Statistically significant differences compared to DMSO-treated cells were determined by a repeated-measures one-way ANOVA with Dunnett’s multiple comparisons test (B–G). Bars indicate mean ± SEM. Points indicate individual replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.
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TYK2 inhibition enhances Treg stability in Th17-polarizing conditions Tregs (CD4 + CD25 hi CD127 lo ) were isolated from peripheral blood of healthy human donors, stimulated with anti-CD3/CD28, and cultured for 7 days in a Th17-polarizing cocktail in the presence of BMS-986202 (B202). (A) Schematic of polarization protocol. (B) The proportion of FOXP3 + cells was assessed after 7 days. Representative plots and quantification are shown ( n = 9). (C) Expression of RORC2, CCR4, and CCR6 was determined after 7 days relative to non-polarized, control Tregs ( n = 3). (D) Methylation of the Treg-specific demethylated region <t>(TSDR)</t> after 7 days of culture ( n = 5). (E) Representative plot and quantification of intracellular IL-17A/F expression after a 4 h stimulation with PMA/ionomycin ( n = 5). (F) IL-17A, IL-17F, and TNF-α concentration in supernatant after a 24 h re-stimulation with anti-CD3/CD28 ( n = 6). (G) Representative figures and quantification of TIGIT and CD226 expression on Tregs on day 7 ( n = 9). Statistically significant differences compared to DMSO-treated cells were determined by a repeated-measures one-way ANOVA with Dunnett’s multiple comparisons test (B–G). Bars indicate mean ± SEM. Points indicate individual replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.
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Pyrosequencing Inc primers for pyrosequencing assays for prame, cldn11, and shox2
TYK2 inhibition enhances Treg stability in Th17-polarizing conditions Tregs (CD4 + CD25 hi CD127 lo ) were isolated from peripheral blood of healthy human donors, stimulated with anti-CD3/CD28, and cultured for 7 days in a Th17-polarizing cocktail in the presence of BMS-986202 (B202). (A) Schematic of polarization protocol. (B) The proportion of FOXP3 + cells was assessed after 7 days. Representative plots and quantification are shown ( n = 9). (C) Expression of RORC2, CCR4, and CCR6 was determined after 7 days relative to non-polarized, control Tregs ( n = 3). (D) Methylation of the Treg-specific demethylated region <t>(TSDR)</t> after 7 days of culture ( n = 5). (E) Representative plot and quantification of intracellular IL-17A/F expression after a 4 h stimulation with PMA/ionomycin ( n = 5). (F) IL-17A, IL-17F, and TNF-α concentration in supernatant after a 24 h re-stimulation with anti-CD3/CD28 ( n = 6). (G) Representative figures and quantification of TIGIT and CD226 expression on Tregs on day 7 ( n = 9). Statistically significant differences compared to DMSO-treated cells were determined by a repeated-measures one-way ANOVA with Dunnett’s multiple comparisons test (B–G). Bars indicate mean ± SEM. Points indicate individual replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.
Primers For Pyrosequencing Assays For Prame, Cldn11, And Shox2, supplied by Pyrosequencing Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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TYK2 inhibition enhances Treg stability in Th17-polarizing conditions Tregs (CD4 + CD25 hi CD127 lo ) were isolated from peripheral blood of healthy human donors, stimulated with anti-CD3/CD28, and cultured for 7 days in a Th17-polarizing cocktail in the presence of BMS-986202 (B202). (A) Schematic of polarization protocol. (B) The proportion of FOXP3 + cells was assessed after 7 days. Representative plots and quantification are shown ( n = 9). (C) Expression of RORC2, CCR4, and CCR6 was determined after 7 days relative to non-polarized, control Tregs ( n = 3). (D) Methylation of the Treg-specific demethylated region <t>(TSDR)</t> after 7 days of culture ( n = 5). (E) Representative plot and quantification of intracellular IL-17A/F expression after a 4 h stimulation with PMA/ionomycin ( n = 5). (F) IL-17A, IL-17F, and TNF-α concentration in supernatant after a 24 h re-stimulation with anti-CD3/CD28 ( n = 6). (G) Representative figures and quantification of TIGIT and CD226 expression on Tregs on day 7 ( n = 9). Statistically significant differences compared to DMSO-treated cells were determined by a repeated-measures one-way ANOVA with Dunnett’s multiple comparisons test (B–G). Bars indicate mean ± SEM. Points indicate individual replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.
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Pyrosequencing Inc internal pyrosequencing primer 5'- ctcatggcactgtactcttct-3
TYK2 inhibition enhances Treg stability in Th17-polarizing conditions Tregs (CD4 + CD25 hi CD127 lo ) were isolated from peripheral blood of healthy human donors, stimulated with anti-CD3/CD28, and cultured for 7 days in a Th17-polarizing cocktail in the presence of BMS-986202 (B202). (A) Schematic of polarization protocol. (B) The proportion of FOXP3 + cells was assessed after 7 days. Representative plots and quantification are shown ( n = 9). (C) Expression of RORC2, CCR4, and CCR6 was determined after 7 days relative to non-polarized, control Tregs ( n = 3). (D) Methylation of the Treg-specific demethylated region <t>(TSDR)</t> after 7 days of culture ( n = 5). (E) Representative plot and quantification of intracellular IL-17A/F expression after a 4 h stimulation with PMA/ionomycin ( n = 5). (F) IL-17A, IL-17F, and TNF-α concentration in supernatant after a 24 h re-stimulation with anti-CD3/CD28 ( n = 6). (G) Representative figures and quantification of TIGIT and CD226 expression on Tregs on day 7 ( n = 9). Statistically significant differences compared to DMSO-treated cells were determined by a repeated-measures one-way ANOVA with Dunnett’s multiple comparisons test (B–G). Bars indicate mean ± SEM. Points indicate individual replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.
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TYK2 inhibition enhances Treg stability in Th17-polarizing conditions Tregs (CD4 + CD25 hi CD127 lo ) were isolated from peripheral blood of healthy human donors, stimulated with anti-CD3/CD28, and cultured for 7 days in a Th17-polarizing cocktail in the presence of BMS-986202 (B202). (A) Schematic of polarization protocol. (B) The proportion of FOXP3 + cells was assessed after 7 days. Representative plots and quantification are shown ( n = 9). (C) Expression of RORC2, CCR4, and CCR6 was determined after 7 days relative to non-polarized, control Tregs ( n = 3). (D) Methylation of the Treg-specific demethylated region <t>(TSDR)</t> after 7 days of culture ( n = 5). (E) Representative plot and quantification of intracellular IL-17A/F expression after a 4 h stimulation with PMA/ionomycin ( n = 5). (F) IL-17A, IL-17F, and TNF-α concentration in supernatant after a 24 h re-stimulation with anti-CD3/CD28 ( n = 6). (G) Representative figures and quantification of TIGIT and CD226 expression on Tregs on day 7 ( n = 9). Statistically significant differences compared to DMSO-treated cells were determined by a repeated-measures one-way ANOVA with Dunnett’s multiple comparisons test (B–G). Bars indicate mean ± SEM. Points indicate individual replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.
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Overview of the data and methods used in this study. a , b , c , d depict the primary analyses, in which a latent variable model was used to determine the association between peripheral <t>TPH2</t> and SLC6A4 methylation and brain levels of 5-HTT and 5-HT 4 . e , f , g describe the sensitivity analyses evaluating the association between SLC6A4 / TPH2 methylation and measures of environmental stress, depressive and anxiety symptoms. h and i show sensitivity analyses used to evaluate potential influence of blood cell proportions in the A-G analyses. Abbreviations: 5-HT 4 : serotonin 4 receptor; 5-HTT: serotonin transporter; CpG: CpG site; TSS: transcription start site; SLE: stressful life events; PBI: parental bonding inventory; BDI: Beck’s depressive index; GAD10: generalized anxiety disorder 10-item; CATS: childhood abuse trauma scale; HAMD6: Hamilton depressive rating scale 6; PSS: perceived stress scale; CpG LV : latent variable including all CpG methylation values
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TYK2 inhibition enhances Treg stability in Th17-polarizing conditions Tregs (CD4 + CD25 hi CD127 lo ) were isolated from peripheral blood of healthy human donors, stimulated with anti-CD3/CD28, and cultured for 7 days in a Th17-polarizing cocktail in the presence of BMS-986202 (B202). (A) Schematic of polarization protocol. (B) The proportion of FOXP3 + cells was assessed after 7 days. Representative plots and quantification are shown ( n = 9). (C) Expression of RORC2, CCR4, and CCR6 was determined after 7 days relative to non-polarized, control Tregs ( n = 3). (D) Methylation of the Treg-specific demethylated region (TSDR) after 7 days of culture ( n = 5). (E) Representative plot and quantification of intracellular IL-17A/F expression after a 4 h stimulation with PMA/ionomycin ( n = 5). (F) IL-17A, IL-17F, and TNF-α concentration in supernatant after a 24 h re-stimulation with anti-CD3/CD28 ( n = 6). (G) Representative figures and quantification of TIGIT and CD226 expression on Tregs on day 7 ( n = 9). Statistically significant differences compared to DMSO-treated cells were determined by a repeated-measures one-way ANOVA with Dunnett’s multiple comparisons test (B–G). Bars indicate mean ± SEM. Points indicate individual replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Journal: Cell Reports Medicine

Article Title: TYK2 inhibition enhances Treg differentiation and function while preventing Th1 and Th17 differentiation

doi: 10.1016/j.xcrm.2025.102303

Figure Lengend Snippet: TYK2 inhibition enhances Treg stability in Th17-polarizing conditions Tregs (CD4 + CD25 hi CD127 lo ) were isolated from peripheral blood of healthy human donors, stimulated with anti-CD3/CD28, and cultured for 7 days in a Th17-polarizing cocktail in the presence of BMS-986202 (B202). (A) Schematic of polarization protocol. (B) The proportion of FOXP3 + cells was assessed after 7 days. Representative plots and quantification are shown ( n = 9). (C) Expression of RORC2, CCR4, and CCR6 was determined after 7 days relative to non-polarized, control Tregs ( n = 3). (D) Methylation of the Treg-specific demethylated region (TSDR) after 7 days of culture ( n = 5). (E) Representative plot and quantification of intracellular IL-17A/F expression after a 4 h stimulation with PMA/ionomycin ( n = 5). (F) IL-17A, IL-17F, and TNF-α concentration in supernatant after a 24 h re-stimulation with anti-CD3/CD28 ( n = 6). (G) Representative figures and quantification of TIGIT and CD226 expression on Tregs on day 7 ( n = 9). Statistically significant differences compared to DMSO-treated cells were determined by a repeated-measures one-way ANOVA with Dunnett’s multiple comparisons test (B–G). Bars indicate mean ± SEM. Points indicate individual replicates. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, and ∗∗∗∗ p < 0.0001.

Article Snippet: TSDR Pyrosequencing primer: AGAAATTTGTGGGGTGGG , Boardman et al. , N/A.

Techniques: Inhibition, Isolation, Cell Culture, Expressing, Control, Methylation, Concentration Assay

KEY RESOURCES TABLE

Journal: Cell reports

Article Title: The exocyst subunit EXOC2 regulates the toxicity of expanded GGGGCC repeats in C9ORF72 -ALS/FTD

doi: 10.1016/j.celrep.2024.114375

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: C9ORF72-V3 pyrosequencing primers, See , Yuva-Aydemir et al. , N/A.

Techniques: Reverse Transcription, Enzyme-linked Immunosorbent Assay, Control, Recombinant, Plasmid Preparation, Software

Overview of the data and methods used in this study. a , b , c , d depict the primary analyses, in which a latent variable model was used to determine the association between peripheral TPH2 and SLC6A4 methylation and brain levels of 5-HTT and 5-HT 4 . e , f , g describe the sensitivity analyses evaluating the association between SLC6A4 / TPH2 methylation and measures of environmental stress, depressive and anxiety symptoms. h and i show sensitivity analyses used to evaluate potential influence of blood cell proportions in the A-G analyses. Abbreviations: 5-HT 4 : serotonin 4 receptor; 5-HTT: serotonin transporter; CpG: CpG site; TSS: transcription start site; SLE: stressful life events; PBI: parental bonding inventory; BDI: Beck’s depressive index; GAD10: generalized anxiety disorder 10-item; CATS: childhood abuse trauma scale; HAMD6: Hamilton depressive rating scale 6; PSS: perceived stress scale; CpG LV : latent variable including all CpG methylation values

Journal: Clinical Epigenetics

Article Title: No association between peripheral serotonin-gene-related DNA methylation and brain serotonin neurotransmission in the healthy and depressed state

doi: 10.1186/s13148-024-01678-y

Figure Lengend Snippet: Overview of the data and methods used in this study. a , b , c , d depict the primary analyses, in which a latent variable model was used to determine the association between peripheral TPH2 and SLC6A4 methylation and brain levels of 5-HTT and 5-HT 4 . e , f , g describe the sensitivity analyses evaluating the association between SLC6A4 / TPH2 methylation and measures of environmental stress, depressive and anxiety symptoms. h and i show sensitivity analyses used to evaluate potential influence of blood cell proportions in the A-G analyses. Abbreviations: 5-HT 4 : serotonin 4 receptor; 5-HTT: serotonin transporter; CpG: CpG site; TSS: transcription start site; SLE: stressful life events; PBI: parental bonding inventory; BDI: Beck’s depressive index; GAD10: generalized anxiety disorder 10-item; CATS: childhood abuse trauma scale; HAMD6: Hamilton depressive rating scale 6; PSS: perceived stress scale; CpG LV : latent variable including all CpG methylation values

Article Snippet: Primers used for pyrosequencing SLC6A4 and TPH2 (XLSX 9 kb) Table S4.

Techniques: Methylation, CpG Methylation Assay

Demographics of the participants included in the primary analyses

Journal: Clinical Epigenetics

Article Title: No association between peripheral serotonin-gene-related DNA methylation and brain serotonin neurotransmission in the healthy and depressed state

doi: 10.1186/s13148-024-01678-y

Figure Lengend Snippet: Demographics of the participants included in the primary analyses

Article Snippet: Primers used for pyrosequencing SLC6A4 and TPH2 (XLSX 9 kb) Table S4.

Techniques: Methylation

Associations between peripheral TPH2 DNA methylation and brain 5-HTT binding a or 5-HT 4 binding b in the healthy cohort. Orange dashed boxes to the left depict the TPH2 CpG sites and the covariates included in the model. Rs45706210 stands for TPH2 rs45706210 G/T SNP. For representation purposes, PET and MR scanner covariates are not reported in the a and b models. Similarly, although included in the 5-HT 4 latent variable model, 5-HTTLPR/rs25531 and BDNF rs6265 genotypes are not reported in ( b ). Scatter plots in c and d depict the relation between TPH2 methylation and 5-HTT LV or 5-HT 4LV in healthy controls ( c , d ), while the relation between TPH2 methylation and 5-HT 4 binding in patients with MDD is showed in ( e )

Journal: Clinical Epigenetics

Article Title: No association between peripheral serotonin-gene-related DNA methylation and brain serotonin neurotransmission in the healthy and depressed state

doi: 10.1186/s13148-024-01678-y

Figure Lengend Snippet: Associations between peripheral TPH2 DNA methylation and brain 5-HTT binding a or 5-HT 4 binding b in the healthy cohort. Orange dashed boxes to the left depict the TPH2 CpG sites and the covariates included in the model. Rs45706210 stands for TPH2 rs45706210 G/T SNP. For representation purposes, PET and MR scanner covariates are not reported in the a and b models. Similarly, although included in the 5-HT 4 latent variable model, 5-HTTLPR/rs25531 and BDNF rs6265 genotypes are not reported in ( b ). Scatter plots in c and d depict the relation between TPH2 methylation and 5-HTT LV or 5-HT 4LV in healthy controls ( c , d ), while the relation between TPH2 methylation and 5-HT 4 binding in patients with MDD is showed in ( e )

Article Snippet: Primers used for pyrosequencing SLC6A4 and TPH2 (XLSX 9 kb) Table S4.

Techniques: DNA Methylation Assay, Binding Assay, Methylation

Results of latent variable models (LVM) evaluating the association between SLC6A4 /  TPH2  methylation and 5-HTT and 5-HT 4 brain binding

Journal: Clinical Epigenetics

Article Title: No association between peripheral serotonin-gene-related DNA methylation and brain serotonin neurotransmission in the healthy and depressed state

doi: 10.1186/s13148-024-01678-y

Figure Lengend Snippet: Results of latent variable models (LVM) evaluating the association between SLC6A4 / TPH2 methylation and 5-HTT and 5-HT 4 brain binding

Article Snippet: Primers used for pyrosequencing SLC6A4 and TPH2 (XLSX 9 kb) Table S4.

Techniques: Methylation

Association between a latent variable including blood cell proportions (CpG LV+cells ) and a latent variable including serotonin transporter (5-HTT) or serotonin 4 receptor (5-HT 4 ) binding

Journal: Clinical Epigenetics

Article Title: No association between peripheral serotonin-gene-related DNA methylation and brain serotonin neurotransmission in the healthy and depressed state

doi: 10.1186/s13148-024-01678-y

Figure Lengend Snippet: Association between a latent variable including blood cell proportions (CpG LV+cells ) and a latent variable including serotonin transporter (5-HTT) or serotonin 4 receptor (5-HT 4 ) binding

Article Snippet: Primers used for pyrosequencing SLC6A4 and TPH2 (XLSX 9 kb) Table S4.

Techniques:

Methodology of studies assessing the gut microbiota composition in cats with suspected or confirmed chronic enteropathies.

Journal: Frontiers in Microbiology

Article Title: Gut microbiota in cats with inflammatory bowel disease and low-grade intestinal T-cell lymphoma

doi: 10.3389/fmicb.2024.1346639

Figure Lengend Snippet: Methodology of studies assessing the gut microbiota composition in cats with suspected or confirmed chronic enteropathies.

Article Snippet: , 15 colony cats with uncharacterized chronic diarrhea , Feces , 16S rRNA gene pyrosequencing Primers for the V1-V2 region.

Techniques: Sequencing, Fluorescence, In Situ Hybridization, Real-time Polymerase Chain Reaction, Bacteria