Fig. 4: MUC1-C and WDR5 are necessary for the activation of the FOS gene.

a Nuclear lysates from BT-549 cells were immunoprecipitated with anti-MUC1-C or a control IgG. Input proteins and the precipitates were immunoblotted with antibodies against the indicated proteins. b RNA-seq was performed in triplicates on BT-549 cells silenced for MUC1 and WDR5. The datasets were analyzed for effects of MUC1-C and WDR5 silencing on repressed and activated genes as depicted by the volcano plots (left) and barplot (right). c Venn Diagram depicting the overlap of 100 downregulated genes and 24 upregulated genes in BT-549 cells silenced for MUC1 and WDR5. d GSEA of the MUC1 (left) and WDR5 (right) RNA-seq datasets using the GOMF DNA BINDING TRANSCRIPTION FACTOR ACTIVITY gene signature. e Lysates from BT-549/tet-MUC1shRNA (left) and BT-549/tet-WDR5shRNA cells (right) treated with vehicle or DOX for 7 days were immunoblotted with antibodies against the indicated proteins. f Genome browser snapshot of ATAC-seq data from the FOS pELS region in BT-549/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days (left). Chromatin was analyzed for accessibility of the pELS by nuclease digestion (right). The results are expressed as % undigested chromatin (mean ± SD and individual values). g BT-549/tet-MUC1shRNA cells were treated with vehicle or DOX for 7 days. Soluble chromatin was precipitated with a control IgG, anti-MUC1-C, anti-NF-κB, anti-SET1A and anti-WDR5 (left) or a control IgG and anti-H3K4me3 (right). h BT-549/tet-WDR5shRNA cells were treated with vehicle or DOX for 7 days. Soluble chromatin was precipitated with anti-WDR5 and anti-H3K4me3 (left). The DNA samples were amplified by qPCR with primers for the FOS pELS. The results (mean ± SD of 3 determinations) are expressed as a percentage of the input DNA for each sample. Chromatin was analyzed for accessibility of the FOS pELS by nuclease digestion (right). The results (mean ± SD of 3 determinations) are expressed as % undigested chromatin.