Fig. 4: MUC1-C regulates expression of the TDP-43 XIST RBP.

a DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days were analyzed for TDP-43 gene transcription (left) and TDP-43 transcripts (right). The results (mean ± SD of three determinations) are expressed as relative levels compared to that obtained for vehicle-treated cells (assigned a value of 1). b DU-145/CshRNA and DU-145/NF-κBshRNA cells were analyzed for TDP-43 gene transcription (left) and TDP-43 transcripts (right). The results (mean ± SD of three determinations) are expressed as relative levels compared to that obtained for vehicle-treated cells (assigned a value of 1). c Schema of the TDP-43 gene with highlighting of the PLS region. Soluble chromatin from DU-145/tet-MUC1shRNA cells treated with vehicle of DOX for 7 days was precipitated with anti-MUC1-C, anti-NF-κB, anti-H3K27ac, H3K4me1 and H3K4me3. The DNA samples were amplified by qPCR with primers for the TDP-43 PLS region. The results (mean ± SD of 3 determinations) are expressed as percent input. d. Genome browser snapshot of ATAC-seq data from the TDP-43 PLS in DU-145/tet-MUC1shRNA cells treated with vehicle or DOX for 7 days (left). Chromatin was analyzed for accessibility by nuclease digestion (right). The results (mean ± SD of three determinations) are expressed as % undigested chromatin. e Lysates from DU-145/CshRNA and DU-145/MUC1shRNA (left) or DU-145/NF-κBshRNA (right) cells were immunoblotted with antibodies against the indicated proteins. f Lysates from DU-145 cells expressing the indicated vectors treated with vehicle or DOX for 7 days were immunoblotted with antibodies against the indicated proteins. g DU-145 cells transfected with a control CsiRNA or TDP-43siRNA were analyzed for expression of the indicated genes. The results (mean ± SD of three determinations) are expressed as relative levels compared to that obtained for CsiRNA cells (assigned a value of 1). Lysates were immunoblotted with antibodies against the indicated proteins (right).