Journal: Journal of Advanced Research

Article Title: The GOLM1-ACLY pathway regulates macrophage-secreted EFEMP1 via H3K27ac modifications to drive tumor progression

doi: 10.1016/j.jare.2025.07.003

Figure Lengend Snippet: Nucleic P-ACLY within Golm1 -knockout macrophage regulates Efemp1 expression through H3K27 acetylation (A) Total histone H3 acetylation levels in Thp1 nc and Thp1 −/− cells determined by Histone H3 acetylation detection Kit. n = 9 replicated wells. (B-C) WB detecting (B) and quantitative analysis (C) of the protein levels of various acetylated histone H3. n = 3 independent experiments. (D) PCA plot of CUT&Tag-seq data of experiments with Thp1 nc and Thp1 −/− cells. n = 3 samples/group. (E) Pie chart showing the distribution of H3K27ac peaks across annotated genomic regions in Thp1 nc and Thp1 −/− cells. (F) Heatmap showing the genomic occupancy of H3K27ac from −3 kb flanking TSS to + 3 kb flanking TES in Thp1 nc or Thp1 −/− cells. The x-axis represents the position of reads relative to the TSS, and the y-axis represents the read density. (G) CUT&Tag-seq volcano plot of different gene peaks between Thp1 nc and Thp1 −/− cells. (H) RNA-seq volcano plot of DEGs between BMDMs wt and BMDMs mko cells. n = 3 mice/group. (I) Overlap of differentially expressed genes identified by RNA-seq and CUT&Tag-seq. Secreted proteins with statistically significant differences are listed below. (J) IGV snapshot of Thp1 nc and Thp1 −/− cells showing the H3K27ac CUT&Tag signals at the Efemp1 genomic loci. (K) H3K27ac occupancy in the genome of Efemp1 analysed by ChIP-qPCR. n = 4 duplicates/group. (L) Relative expression of Efemp1 genes in Thp1 nc and Thp1 −/− cells determined by qPCR. n = 6 duplicates/group. The data are presented as the means ± SEMs. The results presented in (A, C, K, and L) were analyzed via two-tailed Student's t test. ns, not statistically significant. **p < 0.01; ***p < 0.001.

Article Snippet: The Bead-bound cells were resuspended and sequentially incubated with anti-H3K27ac primary antibody (8173S, Cell Signaling Technology, USA) and secondary antibody (TransGen Biotech, China).

Techniques: Knock-Out, Expressing, RNA Sequencing, ChIP-qPCR, Two Tailed Test