ha kdm6b  (Addgene inc)

Journal: Gut microbes

Article Title: Fusobacterium mortiferum and its metabolite 5-aminovaleric acid promote the development of colorectal cancer in obese individuals through Wnt/β-catenin pathway by DKK2.

doi: 10.1080/19490976.2025.2502138

Figure Lengend Snippet: Figure 6. 5-AVA may increase DKK2 methylation levels by inhibiting KDM6B. (a) Analysis of DKK2 CpG methylation and H3K27me3 methylation map. (b) Docking of 5-AVA with KDM6B demethylation targets using MOE software, displaying a 2D planar diagram of the interaction between 5-AVA and its target through amino acid side chains and the backbone receptor. (c) 3D simulation diagram of the

Article Snippet: HCT116 cells were transfected with a plasmid encoding the full-length human KDM6B gene (Addgene #24167) to achieve overexpression.

Techniques: Methylation, CpG Methylation Assay, Software

Journal: The EMBO Journal

Article Title: The growth factor EPIREGULIN promotes basal progenitor cell proliferation in the developing neocortex

doi: 10.1038/s44318-024-00068-7

Figure Lengend Snippet: ( A ) Epigenome editing (EE) employing the catalytic domain of KDM6B (JMJC_6B) fused to nuclease deficient Cas9 (dCas9) in mNSCs. Histone methylation and gene expression were analyzed 2 days post-nucleofection following FACS isolation of GFP-positive cells. ( B ) The location of the gRNAs and primer binding sites (PP, primer pair) for ChIP-qPCR is shown for the Ereg locus. Guide RNAs g Ereg EE1 + 2 and g Ereg EE3 + 4 were co-expressed from one plasmid, respectively. ( C ) Level of H3K27me3 at Hoxb , Eomes, Actb , and Ereg (PP1 to PP3) as determined by ChIP-qPCR in mNSCs. ( D ) Bright-field and GFP fluorescence images of mNSCs 2 days post nucleofection with a dCas9-JMJC_6B-T2A-EGFP-gLacZ plasmid. ( E ) ChIP-qPCR analysis of H3K27me3 around the TSS of Ereg and two unrelated genes ( Hoxb5 , Eomes ) after epigenome editing at the Ereg locus. ( F , G ) Expression of Sox2 and Ereg as determined by RT-qPCR upon epigenome editing using g Ereg EE1 + 2 and g Ereg EE3 + 4. Expression normalized to Gapdh and relative to g LacZ EE. Data information: Scale bar, 100 µm. Bar graphs represent mean values. Error bars represent the SD of three replicates (from two to three independent experiments). One-way ANOVA with Dunnett post hoc test; no statistically significant changes were detected.

Article Snippet: The coding sequence of the catalytic domain of human KDM6B (1025–1680 aa) was obtained from the MSCV_JMJD3 plasmid (Addgene #21212) (Sen et al, ).

Techniques: Methylation, Gene Expression, Isolation, Binding Assay, ChIP-qPCR, Plasmid Preparation, Fluorescence, Expressing, Quantitative RT-PCR