Journal: Oncogenesis
Article Title: MYCN and KAT2A form a feedforward loop to drive an oncogenic transcriptional program in neuroblastoma.
doi: 10.1038/s41389-025-00557-2
Figure Lengend Snippet: Fig. 1 KAT2A is associated with MYCN. A STRING analysis of MYCN protein partners in the “chromatin organization” category shows a significant enrichment of the subunits of the SAGA complex. B The IMR32 whole cell extracts are used for co-IP with anti-MYCN antibody. Western blot assay is used to confirm the pulldown of MYCN cofactors KAT2A and MAX. C IMR32 cell extracts were fractionated through a gel filtration column, and selected fractions were then resolved by SDS-PAGE, and western blotting was performed to identify indicated proteins. D Comparison of KAT2A and KAT2B mRNA levels in the 36 NB cell lines (queried from depmap.org). E Determine KAT2A and KAT2B dependencies in the 39 NB cell lines by analyzing data from genome-wide cancer genetic vulnerability CRISPR screens on DepMap. F Western blot assays show the protein levels of KAT2A and KAT2B in HEK293T cell line and MYCN-amplified NB cell lines. G K-Means clustering of ChIP- seq results of MYCN, histone marks, and KAT2A around MYCN binding sites of NB cell line IMR32 (±3 kb). H GREAT peak distribution shows that approximately 20% of KAT2A peaks derived from IMR32 cells are within 5 kb from TSS (transcription start site), while 80% of KAT2A peaks are 5 kb away from TSS. I GREAT Gene Ontology (GO) analysis indicates that KAT2A binding sites associated genes are enriched in protein synthesis and RNA processing. J ChIPPeakAnno analysis shows the number of common and unique peaks between MYCN and KAT2A. K GREAT Gene Ontology (GO) analysis indicates that KAT2A and MYCN overlapped binding sites associated genes are enriched in protein synthesis and RNA processing. Data information: In panels (D and E), the sample sizes (number of NB cell lines) are displayed on the graph. The middle solid lines indicate the mean. Statistical differences were determined using a two-sided unpaired Student’s t-test.
Article Snippet: The following primary antibodies were used: anti-MYCN antibody (Cat # sc53993), anti-MAX antibody (Cat # sc-197), anti-KAT2B antibody (Cat # sc13124), and anti-GAPDH antibody (Cat # sc-47724) from Santa Cruz Biotechnology, anti-H3K9ac (Cat # 91103) from Active Motif, anti-KAT2A antibody (Cat # ab217876) from Abcam; and anti-acetylated-lysine antibody (Cat# 9441) from Cell Signaling Technology.
Techniques: Co-Immunoprecipitation Assay, Western Blot, SDS Page, Comparison, Genome Wide, CRISPR, ChIP-sequencing, Binding Assay, Derivative Assay
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