rabbit anti ep300  (Cell Signaling Technology Inc)

Journal: Advanced Science

Article Title: KMT2C Loss Promotes NF2‐Wildtype Meningioma Progression and Ferroptosis Sensitivity via Epigenetic Repression of Hippo Signaling

doi: 10.1002/advs.202522756

Figure Lengend Snippet: Loss of KMT2C reprograms the epigenomic landscape. (A) Heatmap showing CUT&Tag‐seq read density for KMT2C in WT IOMM‐Lee and MN‐TY1 cells. (B) Distribution of KMT2C‐binding peaks across different chromatin states. (C) Western blot analysis of histone modifications in KMT2C‐WT and KMT2C‐KO cells. (D) Average signal intensity profiles (top) and corresponding heatmaps (bottom) of CUT&Tag‐seq reads for H3K4me1, H3K27me3, and H3K27ac in WT and KMT2C‐KO IOMM‐Lee cells. (E) Western blot analysis of H3K27ac levels in IOMM‐Lee cells transfected with siRNA targeting CBP or EP300 for 72 h. (C, E) Representative western blots are shown from n = 3 independent biological replicates.

Article Snippet: Antibodies included H3K27ac (CST #8173, 1:100), CBP (GeneTex #GTX101249, 1:100), EP300 (ABclonal #A13016, 1:50).

Techniques: Binding Assay, Western Blot, Transfection

Journal: Advanced Science

Article Title: KMT2C Loss Promotes NF2‐Wildtype Meningioma Progression and Ferroptosis Sensitivity via Epigenetic Repression of Hippo Signaling

doi: 10.1002/advs.202522756

Figure Lengend Snippet: Loss of KMT2C impairs CBP/EP300 function. (A, B) Average signaling intensity curves and heatmap of CUT&Tag‐seq reads for CBP and EP300 in WT and KMT2C‐KO IOMM‐Lee and MN‐TY1 cells. (C) Venn diagram showing genome‐wide overlap among KMT2C, H3K27ac, and CBP/EP300 bound peaks from CUT&Tag‐seq analyses in human meningioma cells. (D–F) Heatmaps displaying differential CBP, EP300, and H3K27ac enrichment at KMT2C co‐occupied regions in WT and KMT2C‐KO IOMM‐Lee cells. (G–I) Aggregate plots showing changes in H3K27ac, CBP and EP300 signal intensity (±3 kb from peak centers) between WT and KMT2C‐KO cells.

Article Snippet: Antibodies included H3K27ac (CST #8173, 1:100), CBP (GeneTex #GTX101249, 1:100), EP300 (ABclonal #A13016, 1:50).

Techniques: Genome Wide

Journal: Advanced Science

Article Title: KMT2C Loss Promotes NF2‐Wildtype Meningioma Progression and Ferroptosis Sensitivity via Epigenetic Repression of Hippo Signaling

doi: 10.1002/advs.202522756

Figure Lengend Snippet: KMT2C regulates NF2 expression via H3K27ac, and NF2 suppresses tumor progression in KMT2C‐deficient meningiomas. (A) Venn diagram showing overlap between genes downregulated upon KMT2C loss (RNA‐seq) and those with reduced H3K27ac enrichment (CUT&Tag‐seq). (B) Genome browser tracks showing H3K4me1, H3K27me3, and H3K27ac peaks at the NF2 locus in WT and KMT2C‐KO IOMM‐Lee cells. (C) CUT&Tag‐qPCR quantification of H3K27ac occupancy at the NF2 promoter in WT and KMT2C‐KO IOMM‐Lee and MN‐TY1 cells (n = 3). (D, E) Western blot and qRT‐PCR analyses showing decreased NF2 protein and mRNA levels in KMT2C‐KO cells (n = 3). (F) Western blot analysis showing reduced NF2 expression following CBP or EP300 knockdown in IOMM‐Lee cells (72 h). (G) Representative IHC staining of KMT2C and NF2 in human meningioma specimens. Scale bar, 50 µm. (H) Western blot validation of NF2 overexpression in WT and KMT2C‐KO IOMM‐Lee cells. (I–K) CCK‐8 and colony formation assays showing that NF2 overexpression suppresses proliferation in WT and KMT2C‐KO IOMM‐Lee cells (n = 3). (L–N) Representative images, tumor growth curves, and weights of subcutaneous xenografts derived from indicated cells (n = 5). (O) Bioluminescence imaging of intracranial tumor burden in nude mice treated with Erastin, at days 7, 14, and 21 post‐implantation. (P) Kaplan–Meier survival analysis of mice bearing indicated tumors (n = 7 per group). (Q) Body weight monitoring during the experimental period (n = 7 per group). (R) IHC staining of Ki‐67 showing reduced proliferation upon Erastin treatment, reversed by NF2 overexpression. Scale bar, 20 µm (n = 5). (D, F, H) Representative western blots are shown from n = 3 independent biological replicates. Data are presented as means ± SD and analyzed by two‐way ANOVA with post hoc test or unpaired two‐tailed Student's t‐test. n.s for p > 0.05, * p < 0.05, ** p < 0.01, ***p < 0.001.

Article Snippet: Antibodies included H3K27ac (CST #8173, 1:100), CBP (GeneTex #GTX101249, 1:100), EP300 (ABclonal #A13016, 1:50).

Techniques: Expressing, RNA Sequencing, Western Blot, Quantitative RT-PCR, Knockdown, Immunohistochemistry, Biomarker Discovery, Over Expression, CCK-8 Assay, Derivative Assay, Imaging, Two Tailed Test