Extended Data Fig. 8: ASXL1 interacts with inflammatory factors including IRAK1-TAK1 complex.
a: Scheme of liquid chromatography-mass spectrometry (LC-MS) with immunoprecipitated-ASXL1 in 293 T cells. 293 T cells were transfected with N-terminus 3x FLAG-tagged ASXL1-WT or N-terminus 3x FLAG-tagged ASXL1-MT. Binding proteins were analysed by LC-MS. b, c: Pathway enrichment of Kyoto Encyclopedia of Genes and Genomes (KEGG) 2021 Human data sets with IP-MS using ASXL1-WT-specific binding proteins in 293 T cells. The top 10 enriched pathways (b) and top 20 genes included in each enriched pathway (c) are shown. For the pathway analysis, proteins that bind to ASXL1-WT 10-fold more than ASXL1-MT were used. Bars represent –log10 (p-value) of interacting proteins that appear in the designated pathway. The terms of the infectious disease gene sets including TLR signalling are coloured red. d: Scheme of endogenous-ASXL1 tagging with 3x-FLAG-P2A-Blasticidin S deaminase (BSD) in THP-1 cells using CRISPR-Cas9. THP-1 cells expressing Cas9 were simultaneously transfected with the appropriate amount of donor plasmid and gRNA plasmid using Lonza 4D-Nucleofector. e: A western blot of ASXL1 with immunoprecipitated samples for the RIME assay. f: The top 20 genes included in top 10 enriched pathway by Enrichr-based KEGG 2021 Human pathway enrichment analysis using 3xFLAG-tagged endogenous ASXL-specific binding proteins in THP-1 cells. The terms of the infectious disease gene sets including TLR signalling are coloured red. The pathway enrichment analyses were performed using Enrichr (https://maayanlab.cloud/Enrichr/)46,47,48.
