Fig 2: Impact of BAZ1B silencing on gene expression in MCF-7 BC cells. A Results of IPA (Ingenuity Pathway Analysis) functional analysis showing statistically significant pathways enriched following BAZ1B silencing in MCF-7 cells for 72 h. B Statistically significant functions highlighted by Gene Set Enrichment Analysis (GSEA) in siRNA-mediated BAZ1B kd cell transcriptome in MCF-7 cells. NES: Negative Normalized Enrichment Score. C Venn diagram showing the number of common transcripts up-regulated (red) and down-regulated (green) following EPZ treatment or siRNA-mediated BAZ1B kd (upper panel) and MI-2 treatment or siRNA-mediated BAZ1B kd (lower panel) in MCF-7 cells. D Heatmap showing common down-regulated (green) and up-regulated (red) transcripts following BAZ1B silencing, EPZ or MI-2 treatments in MCF-7 cells

Fig 3: Effects of BAZ1B silencing in combination with Dot1L or menin blockade in antiestrogen-sensitive and antiestrogen-resistant BC cells. A Proliferation rate assessed by MTT assay in AE-sensitive (MCF7) tamoxifen (MCF7 TAM-R)- and fulvestrant/ICI (MCF7 ICI-R)-resistant BC cells following BAZ1B, Dot1L or menin silencing or combining BAZ1B silencing with EPZ B or MI-136 C treatment. Data are presented as the mean ± SD of multiple determinations from a representative experiment performed in multiple replicates. RT-qPCR showing ESR1 (ERα) mRNA level by combining BAZ1B silencing with EPZ D and MI-136 E pharmacological inhibition in the same experimental condition indicated above. RT-qPCR results represent mean ± SD of triplicate determinations from a representative experiment. Asterisks indicate statistically significant differences (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.005) to CTRL or to single treatment (black bars)

Fig 4: BAZ1B -/- DAPI-dense block relationship to other nuclear structures. (a) DAPI-dense block distribution relative to nucleoli markers MYBBP1A (red, IF) and NOR (red, direct-labeled FISH). (b) DAPI-dense block distribution relative to centromere markers CENP-A (green, IF) and alpha-satellite DNA indicated by a pan-centromeric direct-labeled FISH probe (green). (c) DAPI-dense block distribution relative to PML bodies (green, IF).

Fig 5: Association between BAZ1B, Dot1L and menin and analysis of BAZ1B expression in BC tumors. A Network reprentation interaction between Dot1L, menin, BAZ1B and the proteins of B-WICH (blue) and WINAC (green) complexes found among Dot1L and menin common interactors. B Western blot showing BAZ1B, Dot1L and menin co-immunoprecipitation MCF-7 nuclear extracts. BAZ1B mRNA C and protein D expression levels in luminal-like breast cancer samples from The Cancer Genome Atlas (TCGA) and Clinical Proteomic Tumor Analysis Consortium (CPTAC) datasets analyzed with UALCAN. Kaplan–Meier curves, generated using the Kaplan–Meier Plotter, showing the probability of overall (E; 437 low and 283 high samples, respectively) and relapse-free (F: 457 low and 274 high samples, respectively) survival of ERα + BC patients according to BAZ1B mRNA expression levels