Fig. 4: AIB1 transcriptome and interactome in the endocrine resistant cells acts as a regulator of pro-metastases pathways.

A Scatter plot graph displays % cell viability after 96 h of multiple endocrine resistant cell line models (under estrogen deprived conditions) in the presence of vehicle (DMSO) or AIB1 inhibitor SI-2 (50 nM). All results are mean ± S.E.M., n = 3 and two-sided t tests were used to calculate P values (***P < 0.0001). B Schematic of the RNA-seq experimental design. RNA-seq was carried out in biological triplicate in MCF7 and LetR cells comparing NT (Scramble siRNA) cells versus cells depleted of AIB1 using a SMARTpool ON-TARGETplus siRNA. C Venn diagram displays the number of differentially expressed genes (DEGs) identified with AIB1 knockdown in MCF7 versus LetR cells (logFoldChange ± 1.5, Padj ≤ 0.05). D Graph displays MsigDB Oncogenic signature gene pathways enriched in AIB1-dependent DEGs specific to LetR cells. Also displayed are P values and Z scores for reach significant pathway identified. E Schematic representation of the experimental design for Rapid Immunoprecipitation of Endogenous (RIME) AIB1. AIB1 RIME was conducted in MCF-7 and LetR cells under estrogen deprivation conditions (n = 3 biological replicates). Nonspecific interactions (identified from multiple IgG control replicates) are removed. ET = endocrine therapy. (F) Venn diagram displays AIB1-interacting proteins identified in MCF7 and LetR. G MS-ARC plot shows AIB1-associated proteins specific to LetR cells. The AIB1-associated proteins are clustered according to molecular function, and the length of the line represents the Mascot score. Proteins of interest are highlighted with enlarged bold text. AIB1 is displayed in the MS-ARC plot as the immunoprecipitated protein of interest. (H) Plot of the Gene Ontology (GO) molecular function enrichment test for the LetR only AIB1 associated proteins (hypergeometric test, P-adj ≤ 0.05).