Fig 2: SP1 enhanced the histone acetylation levels in TNPO2 promoter regions by binding to P300. A, The UCSC Genome Bioinformatics Site (http://genome.ucsc.edu/)showed high enrichment of H3K27Ac at the promoter of TNPO2. B and C, Protein expression level of H3, H3K9, H3K14, and H3K27 were detected by Western blot analysis after transfected with siSP1 or transfected with siDYNC1I1 for 48 h. D, Immunoprecipitation assay was performed to test the complex formation of SP1 and P300 physically in HGC-27 cells (*P < 0.05,**P < 0.01; n = 3, student s t test, means ± 95% CI)

Fig 3: CHES1 was acetylated by p300 and deacetylated by HDAC1.A Endogenous IP assay showed the acetylated CHES1 in the presence of pan-HDACs inhibitors. B CoIP assay showed that CHES1 could be acetylated by p300 but not other acetyltransferases. “*” denotes the non-specific band. C CoIP assay showed the disability of catalytic deficient mutants of p300 on the acetylation of CHES1. D, E CoIP assay showed the endogenous interaction between CHES1 and p300 in TNBC. F IP assay showed the effect of HDACs inhibitors on the acetylation levels of CHES1. G The schematic diagram of the main steps for identifying the acetylation lysine sites of CHES1 by mass spectrometry (MS). H Tandem mass spectrum of the peptide (m/z = 491.59) HNLSLNKCFKK containing acetylated K170 and K173 of CHES1. The “ac” above the Lys indicates an acetyl remnant on this Lys. I The schematic representation of the 21 acetylated lysine sites in the protein domains of CHES1. J CoIP assay showed the acetylation levels of wild type CHES1 and its KR mutant.

Fig 4: The hypoxia‐induced p300/XBP1s/Herpud1 axis facilitated RAW264.7 cell M2 polarization. A, Western blot was performed to detect Herpud1 protein levels in the normal, scrambled siRNA and Herpud1 siRNA transfection groups. B, Densitometric quantification showed the ratio of Herpud1 protein to GAPDH protein. ** P <.01 vs the normal group; # P <.05, ## P <.01 vs the hypoxia group. NS indicates no significant difference vs the hypoxia group. RAW264.7 cells were assigned to normal, hypoxia, hypoxia +0.1% DMSO, hypoxia +L002, hypoxia +STF‐083010 and hypoxia +Herpud1 siRNA groups. C, The mRNA levels of the M1‐type macrophage markers IL‐6, iNOS and CD86 were detected by qRT‐PCR. ** P <.01 vs the normal group; # P <.05 vs the hypoxia group. D, The mRNA levels of the M2‐type macrophage markers Arg1 Ym1 and CD206 were detected by qRT‐PCR. ** P <.01, *** P <.005 vs the normal group; # P <.05 vs the hypoxia group. n = 4/group

Fig 5: YY1 coactivator EP300 undergoes LLPS in vitro. (A) EP300 domain structure and predicted IDRs (yellow shades). (B) Droplets of EGFP-EP300-IDRs (top row) and their quantification of numbers and area (bottom row). (C) Droplets of EGFP-EP300-IDR3 and -IDR5 at different protein concentrations. Quantification is at the bottom row. (D and E) EGFP-EP300-IDR3 and -IDR5 droplet formation at different temperatures (D), and in the presence or absence of 5% 1,6-hexanediol (E). In (B–E), representative images are presented, and quantification is shown as mean ± s.e.m. from six droplets’ fields in each group. (F) EGFP-EP300-IDR3 and -IDR5 droplets coalescence. (G) FRAP recovery of EGFP-EP300-IDR3 and -IDR5 droplets. FRAP recovery curves at right are the data of mean ± s.d. (n = 6 droplets). t, half-time of recovery.