modifiedtm histone peptide array  (Active Motif)

Journal: Nucleic Acids Research

Article Title: PHF20 is crucial for epigenetic control of starvation-induced autophagy through enhancer activation

doi: 10.1093/nar/gkac584

Figure Lengend Snippet: PHF20 recognizes H3K36me2 via its Tudor 1 and 2 domains. ( A ) Relative H3K36me2 ChIP-seq peak intensity of PHF20-dependent (states 6 and 8) and PHF20-independent states in chromHMM. ( B ) Screening for histone peptide binding of PHF20 Tudor 1 and 2 domains (Tudor 1&2) with MODified™ Histone Peptide Array. GST-PHF20 Tudor 1&2 construct was detected with GST antibody. Histone peptides with significant binding intensity are indicated with red, yellow, and blue circles. Each dot indicated with the color contains the following histone peptides. red:H3K27me2, yellow:H3K36me2, and blue:H4K20me2. ( C ) Top five histone peptides with the highest binding intensity. Binding intensity was calculated with MODified™ Histone Peptide Array analysis program. ( D ) In vitro peptide binding assay using GST-PHF20 Tudor 1&2 of WT and W97A constructs was performed, followed by immunoblot analysis with anti-GST antibody. ( E ) CUT&RUN assay of Flag-tagged PHF20 constructs on Atg13 DOP region. ( F ) Schematic model for H3K36me2 recognition of PHF20 WT and Tudor domain deletion mutant.

Article Snippet: In vitro histone peptide binding array was performed with MODified™ Histone Peptide Array Kit (13001, Active Motif, Carlsbad, CA, USA) following manufacturer's protocol.

Techniques: ChIP-sequencing, Binding Assay, Modification, Peptide Microarray, Construct, In Vitro, Western Blot, Mutagenesis