Extended Data Fig. 5: Identification of ligustroflavone’s MOA.
From: High-throughput profiling of chemical-induced gene expression across 93,644 perturbations
a, Overlap among the top 30 compounds based on ZhangScore in two cell lines using the JQ1 signature. b and c, Analysis of Pearson correlation (two-sided) of gene expression profiles between JQ1 and ligustroflavone in HEK293T cells (b) and MDA-MB-231 cells (c). d and e, Heat map illustrating the differential expression of genes in HEK293T cells (d) or MDA-MB-231 cells (e) treated with either ligustroflavone or JQ1. f and g, Molecular dynamics simulation of ligustroflavone binding to the BRD4 protein. RMSF of residues respect to their time-averaged positions during the docking process of ligustroflavone and key target BRD4 (f). Percentage occupancy of different amino acid residues of BRD4 with ligustroflavone (g). h, Western blotting showing the BRD4 CETSA binding assay in the presence or absence of 50 μM ligustroflavone at different temperatures. i, The BRD4 band intensities in the CETSA were quantified. Data represent mean ± SEM (n = 3 biological replicates). j and k, Gene Set Enrichment Analysis (GSEA) of HTS2 results from siBRD4 knockdown in MDA-MB-231 cells, showing the normalized enrichment score (NES). The statistical analysis for GSEA used a one-sided permutation-based enrichment test with adjusted P values (p-adjust) to account for multiple comparisons.
