Fig. 2

TRIM33-dependent SPT16 recruitment at macrophage distal regulatory regions. a (Left) Proteins and number of peptides identified following Flag-TRIM33 immunopurification and mass spectrometry in immortalized macrophages (IM). (Right) Immunoprecipitation (IP) of endogenous proteins from IM nuclear extracts followed by immunoblotting (IB). b Overlap between intergenic SPT16 and TRIM33 peaks in BMDM. c Intergenic SPT16 peaks were divided in quantiles based on their tag density. Higher enriched intergenic SPT16 peaks have higher number of peaks that co-localize with TRIM33 peaks. d TRIM33, PU.1 and SPT16 occupancy at the summit of intergenic SPT16/TRIM33 peaks ± 2 kb. e MNase (left) and ATAC (right) profiles at the summit of intergenic SPT16 peaks that co-localized (SPT16/TRIM33) or not (SPT16 only) with TRIM33. f Normalized SPT16 ChIP-seq tag count at most enriched (Top10%) intergenic SPT16/TRIM33 peaks in WT and Trim33^−/− BMDM. ***p < 0.0001, Paired t test. g SPT16 ChIP-seq and ATAC-seq profiles at indicated loci in WT and Trim33^−/− BMDM along with TRIM33 and PU.1 occupancy in WT BMDM. h Immunoblotting of TRIM33, PU.1 and β-ACTIN in NIH3T3 and NIH3T3 transfected with PU.1 (left). PU.1 (middle) and TRIM33 (right) ChIP-qPCR at indicated loci in NIH3T3 and NIH3T3 transfected with PU.1. i SPT16 ChIP-seq (left) and ChIP-qPCR (right) at the Atp1b3 −35 kb region in WT and Trim33^−/− BMDM. Mean ± SEM, n = 3. j SSRP1 and k PU.1 ChIP-qPCR at the Atp1b3 − 35 kb region in WT and Trim33^−/− BMDM. Mean ± SEM, n = 3