Journal: Theranostics

Article Title: High OGT activity is essential for MYC-driven proliferation of prostate cancer cells

doi: 10.7150/thno.30834

Figure Lengend Snippet: Identification of transcription factors that bind to O-GlcNAc marked chromatin. A) Distribution of differently modified histone 3 reads within ±2kb of O-GlcNAc peak. B) . The top 3 motifs enriched for the O-GlcNAc ChIP-seq data showing similarity with known previously identified transcription factor motifs from published datasets. C) O-GlcNAc ChIP-seq consensus overlap with MYC ChIP-seq data reported in this study. D) O-GlcNAc ChIP-seq consensus overlap with MYC ChIP-exo dataset reported previously (GSE73994). E) OSMI-2 disrupts interaction between MYC and HCF-1. Immunoprecipitation (IP) of MYC. IgG denotes negative control. MYC was overexpressed by addition of doxycycline using the LNCaP-MYC cell line for 4 hours either in the presence or absence of 40µM OSMI-2. Experiment was repeated four times. F) Knockout of OGT disrupts the interaction between MYC and HCF-1. Experiment was performed in a mouse embryonic fibroblast cell line that has been genetically engineered to enable removal of OGT gene by addition of 0.5µM Tamoxifen (Tam). After two days of DMSO- or Tam-treatments, cell lysates were prepared and used for immunoprecipitation. Data shown is representative of two biological replicates. G) Overlap of O-GlcNAc (this study), MYC (this study) and HCF-1 (ENCSR000ECH) ChIP-seq data.

Article Snippet: Tamoxifen-inducible OGT knockout mouse embryonic fibroblast (MEF) cell line was obtained from Dr. Natasha Zachara at the CardioPEG CoreC4 (NHLBI P01 HL107153) at Johns Hopkins University School of Medicine .

Techniques: Modification, ChIP-sequencing, Immunoprecipitation, Negative Control, Knock-Out