anti 5 hydroxymethyl cytosine  (Cell Signaling Technology Inc)

Journal: JCI Insight

Article Title: TET2 promotes tumor antigen presentation and T cell IFN- γ , which is enhanced by vitamin C

doi: 10.1172/jci.insight.175098

Figure Lengend Snippet: The expression level of key MHC I antigen presentation genes TAP1 ( A ), TAPBP ( B ), B2M ( C ), and HLA-A ( D ) and their correlation with patient survival predictions were summarized in different human cancer types. Data collected from the Kaplan-Meier Plotter database, with HR indicating hazard ratio and logrank P indicating P value. Human cancer tissue was purchased from Biomax (Methods) and stained for 5hmC, TAP1, TAPBP, B2M, and with DAPI for immunofluorescence imaging in malignant melanoma ( E ) and colon adenocarcinoma ( F ). Scale bars: 100 μm. White boxes show ×20 magnification. The relative density of 5hmC, TAP1, TAPBP, and B2M relative to DAPI was calculated from at least 6 replicates under enlarged fields. Unpaired, 2-tailed Student’s t test with multiple comparisons corrected using Bonferroni’s method was used to determine the P value of positive cells between 5hmC-high samples and 5hmC-low samples. **** P < 0.0001. ( G ) Schematic model of the role of TET2 downstream of IFN-γ signaling and VC in regulating tumor antigen presentation and T cell activity directed to the tumor cell.

Article Snippet: The following antibodies were used in this study: GAPDH (GeneTex, 110118), IgG (Cell Signaling Technology [CST], 3900S), TET1 (Abcam, ab191698), TET2 (CST, 18950S), TET3 (CST, 99980S), NF-κB (p65) (CST, 8242S), STAT1 (CST, 9172S), 5hmC (Active Motif, 39769), 5mC (Active Motif, 61479), 5hmC (CST, 51660S), B2M (CoraLite 488, Proteintech, CL488-13511), TAPBP (LS Bio, LS-B9746-GOS10-100), TAP1 (Proteintech, 11114-1-AP), CD3 Alexa Fluor 700 (Invitrogen, MHCD0324), CD4 PE-Cyanine7 (Invitrogen, MHCD0412), CD8 PE-Alexa Fluor 610 (Invitrogen, MHCD0822), CD69 Super Bright 645 (eBioscience, 64-0691-82), CD25 Bright 436 (eBioscience, 62-0251-82), CD11c FITC (eBioscience, 11-0114-82), and PE-anti–mouse H-2Kb bound to SIINFEKL (Invitrogen, 12-5743-82).

Techniques: Expressing, Immunopeptidomics, Staining, Immunofluorescence, Imaging, Activity Assay

Journal: Journal of Neuroinflammation

Article Title: Tet1 -mediated 5hmC regulates hippocampal neuroinflammation via wnt signaling as a novel mechanism in obstructive sleep apnoea leads to cognitive deficit

doi: 10.1186/s12974-024-03189-2

Figure Lengend Snippet: IH exposure induced the upregulation of TETs and total 5hmC in the hippocampus of mice. ( A ) The illustration of IH modeling. ( B ) Body weight and hippocampal PO 2 . ( C ) BOLD-fMRI analysis of the hippocampus. ( D ) Total amount of 5hmC in the hippocampus using standard DNA dot blot. ( E ) The expression of TET1, TET2, and TET3 in the hippocampus using IHC staining and qPCR. Analysis was performed by Student’s t test. All data are presented as the mean ± standard deviation

Article Snippet: The membrane was blocked with 5% milk and then incubated with primary antibodies against 5hmC (dilution 1: 1000; Cell Signaling Technology) and 5mC (dilution 1:1000; Cell Signaling Technology) antibodies overnight at 4 °C.

Techniques: Dot Blot, Expressing, Immunohistochemistry, Standard Deviation

Journal: Journal of Neuroinflammation

Article Title: Tet1 -mediated 5hmC regulates hippocampal neuroinflammation via wnt signaling as a novel mechanism in obstructive sleep apnoea leads to cognitive deficit

doi: 10.1186/s12974-024-03189-2

Figure Lengend Snippet: IH exposure caused DMG / DDG in the hippocampus of mice. ( A ) Mean normalized counts of 5mC- and 5hmC-enriched regions in the hippocampus. ( B ) ( C ) 5 kb distribution of 5mC and 5hmC peaks upstream and downstream of the TSS. ( D ) GO analysis of the DDGs. ( E ) GO analysis of the DMGs. ( F ) KEGG pathway analysis. ( G ) Cell types enrichment

Article Snippet: The membrane was blocked with 5% milk and then incubated with primary antibodies against 5hmC (dilution 1: 1000; Cell Signaling Technology) and 5mC (dilution 1:1000; Cell Signaling Technology) antibodies overnight at 4 °C.

Techniques:

Journal: Journal of Neuroinflammation

Article Title: Tet1 -mediated 5hmC regulates hippocampal neuroinflammation via wnt signaling as a novel mechanism in obstructive sleep apnoea leads to cognitive deficit

doi: 10.1186/s12974-024-03189-2

Figure Lengend Snippet: IH exposure promoted the binding of 5hmC to Wnt pathway genes in the hippocampi of mice. ( A ) The connection by assimilating RNA-sequencing data into methylation, demethylation and gene expression in promoter regions. ( B ) The expression of Wnt3a , Ccnd2 , Prickle2 , Il1b , Fgf13 , Htr4 , Adcy1 , Snap25 , and Syt1 in the hippocampus using qPCR. ( C ) 5hmC profiles of Wnt3a , Ccnd2 , and Prickle2. ( D ) The demethylation levels of specific regions in Wnt3a , Ccnd2 and Prickle2 using hMeDIP-qPCR. Analysis was performed by Student’s t test. All data are presented as the mean ± standard deviation

Article Snippet: The membrane was blocked with 5% milk and then incubated with primary antibodies against 5hmC (dilution 1: 1000; Cell Signaling Technology) and 5mC (dilution 1:1000; Cell Signaling Technology) antibodies overnight at 4 °C.

Techniques: Binding Assay, RNA Sequencing, Methylation, Gene Expression, Expressing, Standard Deviation

Journal: Journal of Neuroinflammation

Article Title: Tet1 -mediated 5hmC regulates hippocampal neuroinflammation via wnt signaling as a novel mechanism in obstructive sleep apnoea leads to cognitive deficit

doi: 10.1186/s12974-024-03189-2

Figure Lengend Snippet: Correlation analysis of the cognition function and 5hmC demethylation

Article Snippet: The membrane was blocked with 5% milk and then incubated with primary antibodies against 5hmC (dilution 1: 1000; Cell Signaling Technology) and 5mC (dilution 1:1000; Cell Signaling Technology) antibodies overnight at 4 °C.

Techniques:

Journal: Journal of Neuroinflammation

Article Title: Tet1 -mediated 5hmC regulates hippocampal neuroinflammation via wnt signaling as a novel mechanism in obstructive sleep apnoea leads to cognitive deficit

doi: 10.1186/s12974-024-03189-2

Figure Lengend Snippet: Tet1 knockdown alleviated IH-induced hippocampal neuroinflammation in mice. ( A ) The expression of Wnt3a , Ccnd2 , and Prickle2 in the hippocampus using qPCR. Analysis was performed by Student’s t test. All experiments were repeated at least three times. All data are presented as the mean ± standard deviation. ( B ) Total amount of 5hmC in the hippocampus using standard DNA dot blot. ( C ) Differential expression of 23 cytokines was displayed by hierarchical clustering, and 13 of them were significantly upregulated after IH. Analysis was performed by Student’s t test. All data are presented as the mean ± standard deviation

Article Snippet: The membrane was blocked with 5% milk and then incubated with primary antibodies against 5hmC (dilution 1: 1000; Cell Signaling Technology) and 5mC (dilution 1:1000; Cell Signaling Technology) antibodies overnight at 4 °C.

Techniques: Knockdown, Expressing, Standard Deviation, Dot Blot, Quantitative Proteomics