rbm15b  (Proteintech)

Journal: The EMBO Journal

Article Title: RBM15B recognizes H3K79me2 to guide selective m 6 A-modification of mRNA and enhance oncoprotein translation in MLL-r leukemia

doi: 10.1038/s44318-026-00707-1

Figure Lengend Snippet: ( A ) Overlap of m 6 A peaks with H3K79me2 and H3K79me3 marks in the human genome. The histogram shows that the H3K79me2 modification sites had greater overlap with m 6 A peaks than did H3K79me3 sites. Our previous work ChIP-seq data (Wang et al, ; Data ref: Wang et al, ), and our MeRIP-seq and GLORI data obtained from MOLM-13 cells were used for the analyses. ( B ) Distribution of m 6 A modifications to H3K79me2 peaks in MOLM-13 ( MLL-AF9 ) cells. ( C ) Metagene analysis of m 6 A signal density in MLL -r target genes by using MeRIP-seq (left) and GLORI (right). MOLM-13 ( MLL-AF9 ) cells were treated with or without EPZ5676. ( D ) Venn diagrams showing that 65 MLL-fusion protein target genes decreased m 6 A and H3K79me2 modifications in both RBM15B knockdown and EPZ5676-treated cells. ( E ) Genome browser views of H3K79me2 (ChIP-seq) (Perner et al, ; Data ref: Perner et al, ) and m 6 A (MeRIP-seq and GLORI-seq) signals for HOXA9 and MYC in MOLM-13 ( MLL-AF9 ) cells treated with or without EPZ5676 (top) and verified by ChIP- and MeRIP-qPCR, respectively (bottom). The positions of Primer 1 (P1) and Primer 2 (P2) are shown. ( F ) MeRIP-qPCR for MYC and HOXA9 in MOLM-13 ( MLL-AF9 ), MV4-11 ( MLL-AF4 ), HL60 ( AML1-ETO ), and NB4 ( PML-RARα ) cell lines treated with or without EPZ5676. N/D, no signal in the qPCR detection. N = 3 biological replicates. The P values for MYC (left to right): 0.0017, 0.0099, 0.8797, and 0.3151; The P values for MYC (left to right): 0.0007, 0.0363, N/D, and N/D. ( G ) MeRIP-qPCR for MYC in control (NC) and RBM15B-FLAG overexpressing MOLM-13 ( MLL-AF9 ) cells treated with or without EPZ5676. NC (without EPZ5676) vs RBM15B-FLAG (without EPZ5676), P < 0.0001; NC (without EPZ5676) vs NC (with EPZ5676), P = 0.0009; NC (with EPZ5676) vs RBM15B-FLAG (with EPZ5676), P = 0.5739. ( H ) Western blot for RBM15B and METTL3 in subcellular fractions of MOLM-13 ( MLL-AF9 ) cell, including Cyto, CA, and NP. GAPDH and H4 acted as the cytoplasmic and nucleoplasmic controls, respectively. Cyto cytoplasm, CA chromatin-associated fraction; NP: nucleoplasmic fraction. ( I ) Schematic illustration of the RBM15B CUT&Tag followed by high-throughput sequencing analyses. The global DNA-binding sites of RBM15B were detected by anti-RBM15B. Using IgG as a negative control. ( J ) Distribution of m 6 A modifications to the RBM15B DNA-binding peaks. ( K , L ) Meta-analysis plot showing the RBM15B binding profiles across the +5 kb to −5 kb genomic region around the transcription starts site (TSS) of total genes ( K ) and MLL -r target genes ( L ). Profiles of EPZ5676 treatment (red) compared with control (blue) MOLM-13 cells are presented. ( M ) Distribution of RBM15B binding sites with or without EPZ5676 treatment to the nearest H3K79me2 (Wang et al, ; Data ref: Wang et al, ) peaks. ( N ) Genome browser views of RBM15B (CUT&Tag-seq) and IgG signals for MYC in MOLM-13 treated with or without EPZ5676 (left) and verified by ChIP-qPCR (right). The positions of Primer 1(P1) and Primer 2(P2) are shown. P = 0.0162 for P1, P = 0.8164 for P2. Values are mean ± SEM ( n = 3), one-way ANOVA in ( G ), and unpaired two-tailed Student’s t test in ( E , F , N ). NS not significant; * P < 0.05; ** P < 0.01; *** P < 0.001. .

Article Snippet: Rabbit anti-RBM15B , Proteintech , 22249-1-AP.

Techniques: Modification, ChIP-sequencing, Knockdown, Control, Western Blot, Next-Generation Sequencing, Binding Assay, Negative Control, ChIP-qPCR, Two Tailed Test